Vi Sinh Vat Hoc (Duoc Si)

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B Y T
VI SINH VT HC
(DNG CHO O TO DC S I HC) M S: .20.Y.03
Ch o bin son: V KHOA HC V O TO B Y T Ch bin: PGS.TS. CAO VN THU Nhng ngi bin son: ThS. TRN TRNH CNG TS. KIU KHC N
file://C:\Windows\Temp\qeraucnwjb\vi_sinh_vat_hoc.html
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ThS. NGUYN LIN HNG CN. NGUYN L PHI PGS.TS. CAO VN THU Tham gia t chc bn tho: ThS. PH VN THM TS. NGUYN MNH PHA
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Thc hin mt s iu ca Lut Gio dc, B Gio dc & o to v B Y t ban hnh chng trnh khung o to Dc s i hc. B Y t t chc bin son ti liu dy hc cc mn c s v chuyn mn theo chng trnh trn nhm tng bc xy dng b sch t chun chuyn mn trong cng tc o to nhn lc y t. Sch Vi sinh vt hc c bin son da vo chng trnh gio dc ca Trng i hc Dc H Ni trn c s chng trnh khung c ph duyt. Sch c tp th cc nh gio giu kinh nghim ca Trng i hc Dc H Ni bin son theo phng chm: Kin thc c bn, h thng; ni dung chnh xc, khoa hc; cp nht cc tin b khoa hc, k thut hin i v thc tin Vit Nam. Sch Vi sinh vt hc c Hi ng chuyn mn thm nh sch v ti liu dy hc chuyn ngnh Dc s i hc ca B Y t thm nh nm 2008. B Y t quyt nh ban hnh ti liu dy hc t chun chuyn mn ca ngnh trong giai on hin nay. Trong thi gian t 3 n 5 nm, sch phi c chnh l, b sung v cp nht. B Y t chn thnh cm n cc tc gi b nhiu cng sc hon thnh cun sch; cm n GS.TS. Nguyn Vn Thanh, PGS.TS. L Hng Hinh c v phn bin cun sch sm hon thnh, kp thi phc v cho cng tc o to nhn lc y t. Ln u xut bn, chng ti mong nhn c kin ng gp ca ng nghip, cc bn sinh vin v cc c gi ln xut bn sau sch c hon thin hn. V KHOA HC V O TO B Y T
LI NI U
Tp ti liu Vi sinh vt hc ny v c bn bao gm ni dung chnh ca nhng bi ging cho cc lp sinh vin dc c tch lu trong nhiu nm ging dy. Tuy nhin Vi sinh vt hc trong thi i Cng ngh sinh hc ngy nay (thi k hu gii m genom ngi) thay i rt mnh m, nn bn cnh nhng kin thc c bn cn thit cho sinh vin ngnh khoa hc Dc, cc tc gi c gng cp nht nhng kin thc khoa hc mi nht lin quan v Sinh hc, Vi sinh vt y hc, Cng ngh sinh hc v Vi sinh vt hc Dc. Ti liu chia lm 4 phn chnh (khng k phn M u gii thiu tng qut v Vi sinh vt hc) bao gm: Phn th nht: Vi sinh vt hc i cng trnh by nhng kin thc chung v th gii vi sinh vt, nhng mt c li v hi i vi con ngi, hnh thi, sinh l, chuyn ho, di truyn ca vi sinh vt m i din c trng l vi khun. Phn th hai: Nhim trng v min dch trnh by nhng vn v nhim trng min dch: khng nguyn, min dch dch th v min dch t bo, huyt thanh, kim sot iu ho min dch. Phn th ba: Vi sinh vt gy bnh trnh by cc vi khun v vi nm gy bnh chnh c tc hi i vi sc kho con ngi: cc c im vi sinh vt hc, kh nng gy bnh, cc phng php pht hin v tr liu, phng nga. Phn th t: Virus gy bnh trnh by i cng v virus v cc loi virus gy bnh chnh (trong bao gm c cc virus gy bnh thi s hin nay nh HIV, virus cm): phn loi, c im sinh hc, cc phng php pht hin v tr liu, phng nga. Vi tinh thn cu th, khoa hc, khch quan, hy vng rng nhng kin thc c trnh by trong ti
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liu Vi sinh vt hc ny s gp phn to nn mt phn nn tng kin thc cn thit cho cc dc s tng lai. D rt c gng nhng do thi gian c hn nn sch c th cn nhiu khim khuyt. Cc tc gi rt mong nhn c nhng kin ng gp cho sch c hon thin hn trong nhng ln xut bn sau. Xin chn thnh cm n. CC TC GI
DANH MC CH VIT TT
A ADCC ADN (DNA) AIDS ARN (RNA) ADP ASLO AMP APC ATP BCAK BCAT BCG BCTT BHK BM BT BW C CaDPA CD CM CMI CPD 50 CPE CW DAP DIP EAEC ED EDTA EHEC ELISA EMP EPEC ETEC F F+ Adenin Gy c t bo ph thuc khng th Antibody dependent cellular cytotoxicity Acid 2 deoxyribonucleic Hi chng suy gim min dch mc phi (Acquired Immunodeficiency Syndrome) Acid ribonucleic Adenosin diphosphat Anti streptolyzin O Adenosin monophosphat T bo trnh din khng nguyn Antigen presenting cell Adenosin triphosphat Bch cu i kim Bch cu i toan Vaccin phng lao Bch cu ht trung tnh T bo thn chut t Biovar mouse Biovar trachoma Bodet Wassemann Cytosin Calci dipicolinat Phn lp bit ho, Class differenciation Cytoplasmic membrane Mng t bo cht Cell mediated immun Min dch qua trung gian t bo Cytopathic Dose 50% Liu hu hoi 50% s t bo Cytopathic effect Tc dng ph hu t bo Cell wall Thnh t bo Acid diaminopimelic Defective interfering particle To ra ht virus khng hon chnh Enteroadherent E. coli E. coli bm dnh ng rut Entner Doudoroff Pathway Con ng Entner Doudoroff Ethylendiamintetraacetat Entero haemorrhagic E. coli Nhm E. coli gy chy mu ng rut Enzyme Linked Immunosorbent Assay Th nghim hp ph min dch lin kt enzyme Embden Mayerhoff Parnas Pathway Con ng ng phn Entero pathogenic E. coli E. coli gy bnh ng rut Entero toxigenic E. coli E. coli sinh c t rut Fungus Nm Plasmid gii tnh
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FAD FADH2 FHA FMN FTA
Flavin adenin dinucleotid dng oxy ho Flavin adenin dinucleotid dng kh ho
Filamentous hemagglutinin si ngng kt hng cu Flavin mononucleotid Fluorescence Treponema Antibody phn ng min dch hunh quang gin tip G Guanin (hoc Gram()) G Gram(m) G+ (hoc Gram(+)) Gram dng GDP Guanosin dinucleotid GTP Guanosin trinucleotid HAAg Khng nguyn ca HAV HAV Hepatitis A Virus Virus vim gan A HBV Hepatitis B Virus Virus vim gan B HCV Hepatitis C Virus Virus vim gan C HDV Hepatitis D Virus Virus vim gan D HEV Hepatitis E Virus Virus vim gan E hGH Yu t sinh trng ngi HIV Human Immunodeficiency Virus Virus suy gim min dch ngi HLA Human Leucocyte Antigen Khng nguyn b mt bch cu ngi HMP Hexozo monophosphat pathway Con ng hexozo monophosphat HSV Herpes simplex virus ICAM Intercellular Adhere Molecule Phn t kt dnh lin t bo IG Immunoglobulin IL Interleukin IS Insertion sequence KDPG 2keto3deoxy6phosphatgluconat pathway Con ng 2keto3deoxy6phosphatgluconat KKT Khng khng th KN Khng nguyn KN H Khng nguyn lng KN K Khng nguyn v KN O Khng nguyn vch KT Khng th LD50 Median lethal dose Liu cht trung bnh LFA1(Lympho Function Antigen): LGV LPS LT LTA MAC MAF MGG MHC MLD MRSA My NAD+ NADH2 NADP+ Khng nguyn chc nng ca lympho bo Biovar lymphogalunoma venerum Lipopolysaccharid Thermolabile toxin c t rut Acid lipotechoic Membrane attack complex Phc hp tn cng mng Macrophage activating factor Yu t hot ho i thc bo Nhum mu May Grunwald Giemsa Major Histocompatibility Complex Phc hp ho hp m ch yu Minimal Lethal Dose Liu cht ti thiu Methicillin resistant S. aureus Mycobacterium Trc khun lao Nicotin adenosin dinucleotid dng oxy ho Nicotin adenosin dinucleotid dng kh ho Nicotin adenosin trinucleotid dng oxy ho
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NADPH2 NK NST OM OMP PCR PEP PFU PG PHMD PL PT RIA RLB RLO RSV SAD SAT SDA SERODIA SIg T TBC TCA TCBS TCR TPHA TPI U VDRL VK VSV WHO
Nicotin adenosin trinucleotid dng kh ho Natural killer T bo dit t nhin Nhim sc th Outer membrane Lp mng ngoi Outer membrane protein Protein mng ngoi Polymerase chain reaction Phn ng chui trng hp Phosphoenolpyruvat Plaque forming unit n v to vng tan Peptidoglycan Phc hp min dch Phospholipid Pertussis toxin c t ho g Radioimmunoassay Th nghim min dch phng x Rickettsia like Bacteria Vi khun loi Rickettsia Rickettsia like organism C th loi Rickettsia Respiratory Syncytial Virus Virus hp bo h hp Huyt thanh chng bch hu Serum antitetanic Huyt thanh chng un vn Mi trng Sabouraud K thut ngng kt Latex nhanh Surface Immunoglobulin Thymin T bo cht Chu trnh tricarboxylic acid Thiosulfat, Citrate Bile Salts, Saccarose T cell receptor Th th t bo T Treponema pallidum Haemagglutination phn ng ngng kt hng cu th ng Treponema pallidum Immobilisation Bt ng Treponema pallidum Uracil Veneral Disease Research Laboratories Vi khun Vi sinh vt World Health Organisation T chc Y t th gii
M U
Vi sinh vt hc (Microbiology) b mn khoa hc ca khoa hc s sng nghin cu cc VSV (microorganisms), nhng sinh vt vi kch thc khng th quan st c bng mt thng. Hnh thi i th ca cc VSV m chng ta nhn thy c l cc tp on ng o cc VSV. Quan st ring
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r cc c th VSV cn phi s dng cc knh hin vi phng i cc VSV ln. Cc knh hin vi quang hc (knh hin vi phn pha, knh hin vi hunh quang,...) h tr nghin cu cc vi khun, vi nm,... cn nu mun nghin cu su sc hn cn s dng knh hin vi in t. Knh hin vi in t l cng c c lc gip nghin cu c cc virus, cc ht sng b nh nht. Vi sinh y hc (medical microbiology) l ngnh VSV hc chuyn nghin cu cc VSV trong mi quan h vi sc kho ca con ngi (gy bnh, c hi v c li). Vi sinh dc hc (pharmaceutical microbiology) l ngnh Vi sinh vt hc chuyn nghin cu cc VSV trong mi quan h vi thuc, dc cht lin quan n ln men sn xut, bo qun v nh lng. Trong chng trnh o to Dc s ti liu Vi sinh vt hc ny c bin son cho sinh vin i hc Dc nm th 3. Sau khi hc xong gio trnh ny, sinh vin c th: 1. Trnh by c cu to, hnh thi, chc nng ca t bo vi khun, vi nm, cng nh cu to v chc nng ca virus. 2. Trnh by c c im sinh l, sinh ho v di truyn ca VSV. 3. Trnh by c c im, c ch, ng dng ca min dch trong chn on, iu tr v phng nga bnh nhim trng. 4. Trnh by c c im sinh hc, kh nng gy bnh, phng php chn on, phng v iu tr cc bnh nhim khun, nhim nm v virus thng gp.
1. VI NT V LCH S PHT TRIN CA VI SINH VT HC

Vi sinh vt l nhng sinh vt xut hin trc tin trn Tri t, v loi ngi t xa xa tuy cha c bng chng c th nhng thc c s tn ti ca cc VSV v ng dng chng trong i sng nh ln men ru, bia, mui da, lm tng, lm dm, lm sa chua, lm phomat, thuc da,... Trong Kim t thp c Ai Cp cn c c cc hnh v m t cch nu bia cho cc Pharaon. Tuy nhin trong thi k c i cha th cp n hc thuyt khoa hc, m nhng iu cn li ngy nay chng ta tip nhn c l cc quan st, ghi chp, iu ch th cng. n tn th k XVII, nh sng ch H Lan Antonie van Levenhoek (1632 1723) vi sng ch ra knh hin vi quang hc ( phng i khong 270 ln) u tin trn th gii cho php ng quan st c cc vi khun v cc ng vt nguyn sinh m ng gi l nhng ng vt v cng nh b, vi s lng nhiu hn dn s H Lan trong ming. Edward Jenner (1749 1823), bc s ngi Anh l ngi u tin sng ch ra phng php chng m u b cho ngi phng bnh u ma, mt cn bnh ht sc nguy him. V cng chnh bc s Edward Jenner l ngi u tin s dng thut ng vaccin (vaccin, t gc Latin: Vaccinae c ngha l bnh u ma b) trn th gii. Nh bc hc v i ngi Php Luis Pasteur (18221895) m u k nguyn mi nghin cu v sinh l hc VSV. ng c coi l ng t ca vi sinh vt hc vi hng lot pht kin v i. ng chng minh bn cht ca nhiu qu trnh ln men (ancol ethylic, acid lactic, acid acetic,...), thc hin thnh cng th nghim ph nh thuyt t sinh (spontaneousgeneration hypothesis), pht hin nhiu vi khun gy bnh cho ngi v ng vt (vi khun gy bnh than, bo t trng, t cu, lin cu, m no cu (cng vi Chamberland, Roux v Thuillier), pht minh ra vaccin phng chng nhiu cn bnh nguy him (vaccin phng t g, vaccin phng bnh than), v xut sc nht l vaccin phng nga bnh di (ngy 6/7/1885 l ngy m Pasteur dng liu vaccin tr di u tin trn th gii cu mng sng mt b gi ngi Php 9 tui l Joseph Meister b ch di cn). Nh bc hc c Robert Koch (1843 1910) cng tc c lc vi Pasteur. ng pht hin ra vi khun lao, vi khun t v sng to ra phng php phn lp thun khit VSV bng cch nui cy chng trn cc mi trng c (solid medium) v cch nhum mu chng. Hc tr J.R. Petri ca Robert Koch l ngi sng to ra nhng chic hp lng ngy nay mang tn ng. Ngi t nn mng cho Min dch hc (Immunology) l nh khoa hc Nga Ilya Ilitch Metchnikov (1845 1916) vi hc thuyt thc bo ni ting. Nm 1908 ng cng vi Erhlich
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(ngi xut hc thuyt min dch dch th) nhn chung gii thng Nobel. Pht hin s tn ti ca nhng loi VSV cn nh b hn vi khun nhiu c cng u thuc v nh sinh l hc thc vt ngi Nga D.I. Ivanovskii (1864 1920). ng chng minh c s tn ti ca loi sinh vt siu hin vi gy bnh khm thuc l nm 1882. n nm 1897, nh khoa hc ngi H Lan M.W. Beijerinck (1851 1931) t tn cho chng l Virus (ting Latin: Virus c ngha l cht c). Nm 1917 F.H. dHerelle (1873 1949) pht hin ra cc th virus ca vi khun v t tn l thc khun th (bacteriophage). Nm 1928, A. Flemming pht hin tc dng khng sinh ca Penicillium notatum i vi Staphylococcus aureus m ra k nguyn sn xut khng sinh iu tr bnh nhim trng nh ln men VSV (A. Flemming cng vi B.E. Chain v H.W. Florey gii thng Nobel, 1945), cng nh Waksman pht minh ra streptomycin (1944) chng li cc vi khun G (gii thng Nobel, 1952). Cc VSV bin i gen dung hp c cc thng tin m khi thu l hon ton xa l i vi VSV y l thnh tu ca cng ngh sinh hc VSV t nhng nm 70 ca th k XX, l ng gp c tnh bc ngot trong di truyn hc VSV. Vo nhng nm 1983 1984 ca th k XX, nh khoa hc Php Luc Montagnie v bc s ngi M Robert Roll, mt cch c lp, pht hin ra virus HIV gy bnh AIDS hi chng suy gim min dch mc phi ngi ht sc nguy him cho sc kho con ngi. i dch HIV/AIDS ang l him ho to ln i vi con ngi, nht l ngi dn cc nc ang pht trin vi h thng chm sc y t cha hon chnh v t . Bc sang th k XXI nhn loi li phi i mt vi nhng typ bnh mi do virus v vi khun gy ra: bnh SARS hi chng vim phi cp, bnh cm H5N1 lin quan n gia cm, bnh lin cu c ngun gc t ln,... cng nh bnh lao ang quay tr li cng vi HIV/AIDS. Hy vng ca nhn loi ang t vo cc nh nghin cu VSV hc cng nh cc nh khoa hc khc trong tng lai khng xa s to ra c vaccin phng chng hay php tr liu hiu qu nht chng li HIV/AIDS, cng nh cc bnh nu, loi b cc him ho ny nng cao cht lng cuc sng cho con ngi.
2. C IM CHUNG CA VI SINH VT
2.1. Kch thc ca vi sinh vt rt nh b

Kch thc ca cc vi khun, x khun, vi nm rt nh b, thng c o bng n v micromet (1m = 10 3 mm, hay 10 6m); kch thc ca cc virus cn nh hn nhiu nn c o bng nanomet (1nm =103m = 106mm; 1Ao = 107mm = 101nm) mt vi n vi trm nanomet. Kch thc nh b nhng din tch b mt tip xc ca qun th cc t bo VSV li rt ln. V d: mt lng t bo cu khun vi th tch 1cm3 c din tch b mt l 6m2 (6m2/1cm3). y l mt trong nhng iu kin tin quyt rt thun li VSV c th trao i cht mnh m vi mi trng, thc hin cc chc nng sng si ng.
2.2. Vi sinh vt hp th nhiu, chuyn ho nhanh, vng i ngn

Tuy nh b (nu khng ni l nh b nht) nhng VSV li l cc sinh vt c kh nng hp th v chuyn ho cht dinh dng mnh hn hn so vi cc sinh vt khc trong sinh gii nh cc sinh vt bc cao: vi khun lactic (Lactobacillus) trong mt gi c th phn gii mt lng ng lactose nhiu gp 1.000 10.000 ln khi lng c th ca chng. Nu tnh s l O2 m mt mg cht kh t bo sinh vt tiu th trong 1 gi (tc tiu th th tch oxy QO2) th m l hoc m r thc vt l 0,5 4,0, t chc m gan v thn ng vt l 10 20, cn nm men S. cerevisiae l 110, vi khun chi Pseudomonas l 1.200, vi khun chi Azotobacter l 2.000. c im ny cho php VSV ng vai tr to ln trong cc chu k tun hon vt cht trong t nhin, ti to mi trng sng cho sinh gii, cng nh kh nng s dng hu ch cc VSV thch hp trong i sng ca con ngi. Vi nhng kh nng c bit nhng chu k vng i ca VSV li ngn, y cng c th l c tnh ring i mi cht sng ca th gii vi m.
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2.3. Vi sinh vt sinh trng nhanh, pht trin mnh

Nh hp th nhiu v chuyn ho nhanh nn cc VSV c tc sinh trng v pht trin rt mnh m so vi cc sinh vt khc, cc VSV c tc sinh trng v sinh si ny n vt tri hn hn. Vi khun E. coli sng cng sinh trong rut ngi, nu c a vo trong iu kin thch hp v sau thi gian tim pht, trung bnh c sau 12 20 pht li sinh ra mt th h mi (s t bo hoc sinh khi kh tng gp i). Tnh theo l thuyt (vi thi gian th h 20 pht) th sau 1 ngy m (phn chia 72 ln) t mt t bo ban u c th sn sinh ra lng VSV c sinh khi (biomass) nng ti 4722 tn! Tuy nhin trong thc t, lng vi khun ny ch t n 1010 1012 t bo trong 1ml dch nui cy m thi. Nm men S. cerevisiae, vi thi gian th h 110 120 pht, khi ln men SCP (single cell protein) cho tc to sinh khi cao hn ca b gp 100.000 ln. Thi gian th h ca to Chlorella l 7 gi, ca vi khun lam Nostoc l 23 gi.
2.4. Vi sinh vt thch ng mnh v d pht sinh bin d

Nng lc thch nghi ca VSV vt xa ca ng vt v thc vt. V kch thc nh buc VSV phi un theo s ng nh ca mi trng tn ti v pht trin, nhng cng chnh v th m tri qua hng trm triu nm tin ho, VSV to ra nhng c ch iu chnh chuyn ho thch hp thch nghi vi iu kin sng bt li ca mi trng. Lng enzym thch ng c trong cc t bo VSV chim ti khong 10% protein ni chung chng t kh nng bin i vt cht, ci to mi trng thch nghi cao ca chng. Do c nhiu kh nng bin i thch nghi, cc VSV c th sng, pht trin c ngay c trong cc iu kin rt khc nghit. Mt s loi c th sng c trong cc iu kin hon ton khng c oxy t do (VSV k kh), mt s loi (nm) c th tn ti v pht trin thnh vng dy trong cc b ngm xc c nng phenol cao. Mt s VSV c th thch nghi vi nhit trn 70OC, mt s khc li c th tn ti nng mui n 32%. Vi khun Thiobacillus thioxidans c th sinh trng pH 0,5, cn Thiobacillus denitrificans li thch nghi vi pH 10,7, Micrococcus radiodurans chu c bc x cng cao. ni su nht trong i dng 11.034m (p lc ln hn 1.104 atm) vn c VSV sinh sng, ch yu l cc vi khun lu hunh. VSV cng d pht sinh cc bin d, y cng l c tnh t nhin ca cht liu di truyn trong sinh gii. Hnh thc bin d hay gp nht l t bin gen (gene mutation). Nhng t bin ny thng xy ra vi tn sut 1010 105 lm cho VSV c nhng thay i v hnh thi, cu to, kiu chuyn ho vt cht, cu trc khng nguyn, kh nng khng thuc,... Bn cnh nhng t bin c li c s dng cng c nhng t bin c hi cn gim thiu hoc chng li.
2.5. Vi sinh vt phn b rng, chng loi nhiu

VSV phn b khp mi ni trn Tri t. VSV c mt trong t; trong nc; trong khng kh; trn v trong cc vt dng; trn v trong c th ca nhiu loi sinh vt. Nh vy c th tm thy cc VSV mi ni. V chng loi, cc VSV cng ht sc phong ph: ngy nay con ngi bit n trn trm nghn loi VSV. Tuy nhin theo quan nim ca nhiu nh khoa hc, chng ta mi ch bit c khong 1 2% s loi VSV c trong t nhin m thi v kh nng phn lp, pht hin c cc VSV mi trong tng lai vi nhng tnh trng mi n nay cha bit vn c xc sut rt ln.
3. V TR CA VI SINH VT TRONG SINH GII

Nghin cu th gii hu sinh, cc nh khoa hc c nhiu c gng phn chia sinh gii ra thnh mt s gii d h thng ho, trong h thng phn gii ca Whittaker (1969) c chp nhn rng ri bao gm 5 gii: Monera, Protista, Plantae, Fungi v Animalia. Cc VSV chim hon ton gii Monera v mt phn gii Fungi. Tuy nhin trong h thng phn gii ny cn phi b sung cc virus vo thnh mt lp ca gii Monera. C h thng phn gii chia th gii hu sinh thnh 3
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nhm, 6 gii, cng kh thng dng. Tn khoa hc ca VSV c gi theo ting Latin bao gm 2 t: t u ch tn chi (Genus) c vit hoa, t sau ch tn loi. Mi gn y, trong ti liu chuyn ngnh, tn khoa hc ca mi loi VSV c vit nghing. V d: Bacillus cereus, Bacillus subtilis, Clostridium acetobutilicum, Diplococcus pneumoniae, Escherichia coli, Haemophyllus influenzae, Micromonospora purpurea var. nigrecens, Nocardia mediteranei, Penicillium chrysogenum, Penicillium notatum, Penicillium maneffei, Pseudomonas aeruginosa, Streptomyces aureofaciens, Streptomyces fradiae, Streptomyces griseus, Streptomyces kanamiceticus, Streptomyces noursei,... Vit tt tn khoa hc c vit t u bng ch ci u vit hoa, t th hai vit nguyn gc. V d Escherichia coli vit tt l E. coli.
4. PHN B VI SINH VT TRONG T NHIN

Nh cp trn, VSV c phn b ht sc rng ri trong t nhin. Mi quan h gia cc VSV vi nhau v vi cc sinh vt khc to nn khu h sinh thi. VSV ng vai tr ht sc quan trng trong tun hon vt cht ca t nhin, nhiu chng loi VSV rt c ch cho i sng v kinh t, tuy nhin cng c nhiu loi VSV c hi cho con ngi khi l nguyn nhn gy ra cc bnh truyn nhim nguy him.
4.1. Vi sinh vt trong t v trong nc

Nc c th l ni cc VSV u tin xut hin nh l sinh vt sng nguyn thu c i u tin. Ngy nay t v nc l mi trng t nhin thch hp cho cc loi VSV, nn s lng v chng loi cc VSV trong cc mi trng ny l ht sc phong ph, v trong t l phong ph nht. Trong 1 gam t ly tng canh tc thng c 1 22 t vi khun, 0,5 14 triu x khun, 3 50 triu vi nm,... VSV trong nc c mi quan h mt thit v l hm s ca VSV trong t xung quanh. S lng v chng loi VSV ph thuc vo cht lng ngun nc. Nc trong khe sui thin nhin nguyn sinh cha t VSV. Nc bin i dng cng c cc loi VSV chu c mui sinh si pht trin. Ngun nc thi thnh ph b nhim bn c cha nhiu VSV.
4.2. Vi sinh vt trong khng kh

Khng kh khng phi l mi trng thun li cho VSV, nhng li l ng truyn bnh nguy him gy ra cc bnh ng h hp. Trung bnh c khong 10.000 ht trong 1m3 khng kh, trong khong 4.000 4.500 bo t, t bo vi khun Gram(+) v Gram(), nh bo t nm. VSV trong khng kh thng t bi, t c gi cun vo hay pht tn t cc c quan sinh sn mang nh bo t. cao khong 84km trong khng kh ngi ta vn pht hin thy c VSV. Cc vi khun gy bnh ng h hp c th t cc ht nc bt nh ca bnh nhn khi giao tip m truyn vo ng h hp ca ngi lnh gy bnh. Cc cht thi ca chim di c mang mm bnh cng c th l ngun ly cc bnh c quy m khu vc v ton cu ngy nay, v d, bnh cm gia cm,... Cc VSV thng gp trong khng kh l vi khun lao, vi khun bch hu, lin cu, t cu, ph cu v cc virus thuc nhm Myxo.
4.3. Vi sinh vt c th ngi lnh

Trn b mt da v trong cc khoang ca c th c kh nhiu chng loi VSV k sinh. Phn ln chng l cc VSV lnh tnh khng gy bnh, nhiu loi l hu ch. Cc VSV ng tiu ho tham gia vo cc qu trnh hp th chuyn ho ca c th, cn cc VSV khc c tr trn da v trong cc khoang c th to nn mt khu h sinh thi n nh c vai tr bo v c th. Nu nh khu h sinh thi ny b ri lon, chng hn khi dng khng sinh ng rut ko di s gy ra chng lon khun ng rut. Khu h sinh thi cc VSV n nh cn c kh nng ngn chn cc VSV gy bnh thm nhp v chim ch trong cc b phn ca c th. K sinh trn da ch yu l cc vi khun Gram(+) nh cc loi cu khun, cc Corynebacterium (C. pseudodiphterium v C. xeroxis) khng c c lc.
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Trong ng tiu ho ming c nhiu VSV pht trin. l cc loi cu khun, trc khun G , vi khun k kh v mt s xon khun. Trong h thng d dy rut c nhiu loi VSV c tr. Do pH acid trong d dy, v pH kim cng nh nhiu loi enzym trong rut non m nhng ni ny c t VSV. Ngc li, rut gi c s lng VSV rt ln, chim ti 1/3 khi lng phn kh v ch yu l cc VSV k kh, nhiu nht l vi khun Bacteriodes fragilis vi s lng ln n 1010 1011 t bo/g phn, gp 100 1.000 ln s lng vi khun E. coli. E. coli chim a s trong cc VSV hiu kh, vi t l chim khong 1%. Cc VSV trong ng tiu ho ng vai tr quan trng trong tiu ho trit cc cht cn b thc n cng nh cung cp cho c th mt s nhm B, vitamin K, ngoi ra cn bo v c th theo c ch chim ch trc. Trong ng h hp, hc mi thng c cc loi vi khun k sinh nh t cu vng, vi khun gi bch hu, phn su hn c th c cc loi lin cu, ph cu, Neisseria, Haemophillus influenzae. Trong kh qun v ph qun hu nh khng c VSV k sinh.
5. CC SN PHM DC DNG C NGUN GC T VI SINH VT

Trong t nhin c rt nhiu chng loi VSV c ch c kh nng sinh tng hp ra cc sinh dc cht, v ang c con ngi khai thc, ng dng trong sn xut dc phm bo v sc kho con ngi v trong nhiu ngnh kinh t khc nh th y, chn nui, trng trt, bo qun thc phm,... y ch xin s b lit k mt s lp sn phm quan trng c ng dng trong Y Dc hc, c ngun gc l sn phm trao i cht ca VSV. Cc cht khng sinh. Cht thay th huyt tng. Cc acid hu c, cc acid amin, cc 5nucleotid, cc vitamin v cc enzym. Cc sn phm bin i sinh hc (bioconversion products). Cc sn phm min dch. Cc sn phm cng ngh ti t hp ADN. Vi s pht trin mnh m ca khoa hc v k thut hy vng rng trong tng lai VSV hc vi tim nng v tn s c nhng thnh tu to ln cng ng gp vo s pht trin chung ca nhn loi.
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PHN MT
Chng 1
HNH THI V CU TO T BO CC VI SINH VT PROCARYOTA
Nh nu trn, VSV l nhng sinh vt kch thc rt nh b, bao gm VSV nhn nguyn thu (procaryota), VSV nhn tht (eucaryota) v cc virus. Trong chng ny chng ta ch cp n cc VSV procaryota. VSV tin nhn bao gm vi khun tht (Eubacteria) v vi khun lam (Cyanobacteria). Vi khun tht bao gm vi khun (Bacteria), x khun (Actinomycetes), v vi khun nguyn thu (Chlamidia, Mycoplasma, Rickettsia).
1. VI KHUN (BACTERIA)
1.1. Hnh thi v kch thc vi khun

Vi khun c hnh thi, kch thc a dng, khc nhau v c sp xp theo cch thc khc nhau. Nhng dng hnh thi ch yu ca vi khun l dng hnh cu, dng hnh que v dng xon, ngoi ra c dng c cung v dng si. ng knh ca a s vi khun thay i trong khong t 0,2 2,0m, chiu di nhn chung t khong 1,5 8,0m.
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1.1.1. Cc cu khun (Cocci) Cc vi khun dng hnh cu rt a dng, tu theo cch thc lin kt cc t bo, mt giao tip m phn ra thnh cc chi sau: n cu khun: y l cc cu khun pht trin ring r, khng c s gn kt t bo: Monococcus Micrococcus, v d: Micrococcus pyogenes. T cu khun (Staphylococcus): Cc cu khun lin kt vi nhau thnh tp on nh chm nho. c trng c cc loi Staphylococcus aureus, Staphylococcus epidermidis v Staphylococcus saprophyticus,... Song cu khun: Cc cu khun lin kt vi nhau to thnh cp sng i. Tu thuc vo lin kt theo trc t bo to thnh cc Diplococcus Gram(+), v d: D. pneumoniae; hay lin kt mt bn m to thnh chi Neisseria Gram(), v d: Neisseria meningitidis, N. gonorrhoeae,... Trong s cc song cu khun (nh cc v d nu trn) mt s gy bnh nguy him cho ngi. T cu khun (Tetracoccus): Cc cu khun lin kt vi nhau to thnh tp hp 4 cu khun. Lin cu khun (Streptococcus): Cc cu khun lin kt vi nhau to thnh chui: Streptococcus cremoris, Streptococcus faecalis, Streptococcus lactis, Streptococcus pneumoniae, Streptococcus pyogenes, Streptococcus viridans,... 1.1.2. Vi khun hnh que (Bacterium s nhiu Bacteria) Trc khun l nhm cc vi khun ht sc a dng, chia lm nhiu b vi s lng ln cc chi, loi bao gm cc trc khun i kh khng sinh bo t hoc sinh bo t, trc khun hiu k kh tu tin, trc khun k kh bt buc, sinh bo t hoc khng sinh bo t, bao gm cc loi sau: Trc khun Gram() khng sinh bo t: Escherichia coli (E. coli), E. vulneris, Haemophylus influenzae, Klebsiella pneumoniae, Proteus mirabilis, Proteus vulgaris, Pseudomonas aeruginosa, Pseudomonas denitrificans, Pseudomonas mallai, Pseudomonas pseudomallei, Salmonella typhii, Salmonella typhimurium, Shigella flexneri,... Trc khun hiu kh sinh bo t (nha bo): Cc Bacillus, v d: Bacillus anthracis, Bacillus cereus, Bacillus pumillus, Bacillus subtilis, Bacillus thuriensis,... C chi Bacillus ny c B. anthracis gy bnh cho ngi v ng vt. Trc khun k kh khng sinh bo t: Lactobacillus acidophylus, Lactobacillus bulgaricus, Lactobacillus casei, Lactobacillus delbruecki. Cc Lactobacillus l cc vi khun c ch, cng nh Propionibacterium shermani. Trc khun k kh tu tin: Klebsiella pneumoniae. Trc khun k kh sinh bo t (nha bo): Cc Clostridium, v d: Clostridiun acetobutilicum, Clostridium botulinum, Clostridium butiricum, Clostridium tetani,... Trong s ny c Clostridium tetani, gy bnh un vn, Clostridium botulinum gy ng c thc n,... 1.1.3. Xon khun Trong s cc xon khun th phy khun c th coi l dng trung gian gia trc khun v xon khun thc th. Phy khun (Vibrio): Vi khun dng du phy, loi gy bnh l Vibrio cholera, Vibrio parahaemolyticus, Vibrio minicus, Vibrio vulnificus,... Dng xon tha (Spirillum): T bo dng hnh sin gin, v d: Spirillum rubrum, Spirillum serpens. Dng xon kht (Spirochatales): T bo c dng xon l xo, in hnh l cc Borelia recurrentis, Leptospira australis, Leptospira ictero haemorrhagiae, Leptospira interrogans, Leptospira sejroe, Treponema pallidum,... T bo vi khun c kch thc rt nh b. ng knh ca phn ln vi khun nm trong khong 0,2 2,0m, chiu di t bo vo khong 1,5 8,0m, v khi lng rt nh: 1 t t bo E. coli ch nng 1mg (hnh 1.1).
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Hnh 1.1. Hnh thi cc dng t bo vi khun A. n cu; B. T cu; C, D. Song cu; E. T cu; F. Lin cu; G, H, I.Trc khun; K. Phy khun; L, M. Xon khun.
1.2. Cu to
Vi khun thuc nhm VSV procaryota nn cu to t bo c c th ring gm mt s b phn chnh nh sau: thnh t bo, mng t bo cht, vng nhn (th nhn), t bo cht, v nhy, lng, pili, bo t (nha bo),... (hnh 1.2).
Hnh 1.2. S cu to t bo vi khun
Theo tnh cht bt mu khi nhum mu Gram, ngi ta chia cc vi khun thnh hai nhm: vi khun Gram dng (Gram(+)) v vi khun Gram(m) (Gram()). S khc bit hai nhm vi khun ny c ngun gc t nhng khc bit c bn trong thnh phn ho hc v cu to thnh t bo ca chng. 1.2.1. Thnh t bo (vch t bocell wall)
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D rt b nh, nhng mi t bo vi khun l mt c th sng c lp, tnh c lp ny quy nh cu to ca lp ngoi cng thnh t bo vi khun. Thnh t bo l lp cu trc lin thng khp kn ngoi cng ca vi khun, c bn chc vi chc nng duy tr ngoi hnh t bo, bo v t bo chng li nhng iu kin ngoi cnh bt li. Nng cc cht tan ca t bo cht bn trong t bo thng cao hn bn ngoi, nu khng c thnh t bo vng chc, t bo s b n bi p sut thm thu. Sau khi lm co nguyn sinh cht ri quan st di knh hin vi in t, c th nghin cu c thnh t bo. C mt s dng vi khun khng c vch t bo, chng hn nh sau: th nguyn sinh (protoplast G+), th cu (t bo trn sphaeroplast G), vi khun dng L (L formes hay Lphase variants Streptobacillus moniliformis) v Micoplasma. Cc chc nng ch yu ca thnh t bo vi khun: Duy tr ngoi hnh t bo v gip khng vi tc ng t bn ngoi (v d vi khun Gram(+) chu c p sut thm thu ti 10 15atm, vi khun Gram() chu c p sut thm thu ti 5 10 atm). Ngn tr s xm nhp ca mt s cht c hi (v d thnh t bo vi khun Gram() c th cn tr s xm nhp ca mt s khng sinh). H tr chuyn ng ca lng. Tham gia v kt thc qu trnh phn bo. Cha cc c trng KN ca vi khun. Thnh t bo vi khun l h thng sng c cu trc phc tp. Thnh t bo vi khun Gram(+) khc xa so vi thnh t bo vi khun Gram(). Cc thnh phn cu to ca thnh t bo vi khun Gram(+) v Gram() cng rt khc nhau (bng 1.1).
Thnh t bo l polyme glycopeptid bao gm peptidoglycan v mucopeptid, hay cn gi murein to thnh polyme xp khng tan trong cc dung mi thng, kh chc v bn vng, bao quanh t bo nh mt mng li. Thnh phn c bn ca peptidoglycan bao gm 3 cu t: Nacetylglucosamin, acid Nacetylmuramic v tetrapeptid cha c L v D acid amin. Hai monome Nacetylglucosamin v acid Nacetylmuramic trng hp xen k vi nhau to thnh chui polyme. Tetrapeptid trn mi chui peptidoglycan lin kt vi tetrapeptid trn cc chui khc to thnh lp, v tetrapeptid trn cc lp khc nhau lin kt cho vi nhau to thnh mng li khng gian a lp vng chc bao quanh ngoi mng nguyn sinh cht. ng thi cc thnh phn ca li khng gian lin tc c m ra monome mi c th gn thm vo, nh m t bo c th sinh trng, phn chia c. Thnh t bo ca vi khun Gram(+) l h thng mng li peptidoglycan lp th ba chiu, l polyme gn kt a chiu vng chc (hnh 1.3). Thm vo vi lp peptidoglycan cc vi khun Gram (+) cn c acid teichoic. Acid teichoic l polyme ca Dalanin v glycerol phosphat lin kt vi peptidoglycan hoc mng t bo cht. Loi acid teichoic lin kt vi mng t bo cht c gi l acid lipoteichoic (hnh1.5). Do tch in m, acid teichoic gip vn chuyn cc ion dng vo, ra t bo v cng l d tr phosphat. Ngoi ra, acid teichoic cn c coi l th th hp th c bit i vi mt s thc khun th (Bacteriophage). Bao bn ngoi peptidoglycan c th l polysaccharid hoc polypeptid, thng ng vai tr l khng nguyn thn c hiu.
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Hnh 1.3. S cu trc cc lp bn ngoi t bo vi khun A. Thnh t bo Gram(+); LTA: acid lipoteichoic; B. Thnh t bo Gram().
Khc vi cu trc peptidoglycan lin thng lp th a chiu ca thnh t bo ca vi khun Gram (+), thnh t bo vi khun Gram() c cu trc hai lp r rt (hnh 1.3): lp peptidoglycan mng bn trong v cch mt lp khng gian chu cht l lp mng ngoi (outer membrane OM) (hnh 1.3, 1.4). Lp mng ngoi c cu to 2 lp gm mt lp phospholipid bn trong kt hp vi mt lp lipopolysaccharid (LPS) bn ngoi an xen vi cc phn t protein v lipoprotein. Lp lipopolysaccharid c cu to bi 3 thnh phn chnh (hnh 1.6) bao gm: Lipid A c cu thnh bi 5 chui di acid bo v 2 phn t Nacetylglucosamin. Polysaccharid li cha: + Li trong gm: + 3 phn t KDO (acid ketodeoxyoctulosonic). + 2 phn t Hep (LglycerinDheptose).
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+ Li ngoi gm: + Cc phn t hexose (bao gm glucosamin, galactose v glucose). Khng nguyn O (KNO) l phn polysaccharid vn ra ngoi mng vo mi trng, gm cc phn t hexose (bao gm galactose, rhamnose, mannose v abequose). Lipid A l ni c t ca vi khun, c th gy st, tiu chy, ph hu hng cu v dn n sock nguy him. KNO quyt nh mt s c tnh khng nguyn ca vi khun c cha LPS v l th th ca bacteriophage. Mng ngoi (OM) ca thnh t bo vi khun Gram() cn cha mt s loi protein, nh protein nn, protein mng ngoi, lipoprotein. Protein nn (matrix protein) nh porin vi khun E. coli l loi protein l, nm xuyn sut qua mng ngoi v cho php mt s loi phn t i qua nh ng, acid amin, dipeptid, tripeptid v cc ion. Protein mng ngoi (Outer Membrane Protein, OMP) l protein vn chuyn c kh nng vn chuyn mt s phn t kh ln ra ngoi, nh vitamin B12, nucleotid, fericrom, enterokelin. Lipoprotein: C nhiu chng loi lipoprotein vi phn t lng 7.200Da, c chc nng lin kt lp peptidoglycan bn trong vi lp mng ngoi.
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Lp khng gian chu cht l lp lu khng nm gia lp peptidoglycan mng v lp mng ngoi thnh t bo vi khun G v cng c phn tip gip gia mng t bo cht v thnh t bo ca c vi khun G+ v G. Trong lp lu khng ny c nhiu loi enzym, chng hn protease, nuclease, protein vn chuyn qua mng, protein th th bacteriophage.
Nhum Gram: Nhum Gram l phng php nhum mu thch hp quan st cc t bo vi khun. Trong phng php Gram, t bo vi khun trc ht c x l vi thuc nhum tm Gentian carboxylic ri vi dung dch lugol, dn n vic to thnh phc cht tm Gentianiod bn trong thnh t bo. Khi vi khun Gram () b ty cn, lipoid ca lp mng ngoi b ho tan lm tng tnh thm ca mng v lm ra tri mt phc cht tm Gentianiod, lm cho vi khun tr li khng mu. Khi nhum b sung fuchsin, t bo s bt mu vi thuc nhum ny (mu vng hay ta). vi khun Gram(+), cn lm cho cc mao qun trong peptidoglycan co li, do phc cht tm Gentian iod b gi li trong thnh t bo v vi khun ch bt mu tm Gentian.
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Thnh ca t bo vi khun l ni tc dng ca nhm khng sinh quan trng (lactam), ng thi cng l ni tc dng ca lysozym, m ch u c lin quan n peptidoglycan (ch tc dng ca khng sinh l peptidoglycantranspeptidase v Dalanincarboxypeptidase). Chnh v th khng sinh penicilin v lysozym tc dng mnh ch yu ln vi khun Gram(+) v tc dng yu ln vi khun Gram(). 1.2.2. Mng t bo cht (cytoplasmic membraneCM) Mng t bo cht l lp mng nm st pha trong thnh t bo, dy khong 4 5nm, cha khong 60% protein v 40% phospholipid. Phospholipid (PL) to thnh mng 2 lp lin thng, cn cc protein c th nm pha trong, pha ngoi hay xuyn qua mng. Cc phn t PL cha u tch in phn cc (u phosphat) v ui khng phn cc (hydrocarbon). u phn cc (ho nc) tan trong nc quay ra pha ngoi mng, hoc hng vo t bo cht bn trong, hoc hng ra pha thnh t bo. Cc PL c trong mng lm mng chy lng v nh vy cc protein c vn ng tnh tnh trong mng cao. y l th gi lng ng hc cn thit cho vic m bo chc nng ca mng cn gi l m hnh khm lng (hnh 1.7).
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Hnh 1.7. Cu trc ca mng t bo cht
Hu ht mng t bo cht ca vi khun khng cha sterol, nh cholesterol, do khng vng chc nh CM ca cc t bo nhn tht. Mycoplasma l h vi khun nguyn thu khng c thnh t bo, nhng CM ca chng c cha sterol, do kh vng chc. Cc chc nng c bn ca CM gm: CM l hng ro i vi a s cc phn t tan trong nc v c tnh chn lc cao hn so vi thnh t bo. Tuy vy CM c cha cc protein vn chuyn c bit cn c gi l permease, c th vn chuyn c cc cht phn t nh vo t bo theo c ch th ng hay ch ng. Cc permease c hiu vi tng cht hoc nhm cht v ATP. CM vi khun l ni m t bo thc hin cc chc nng h hp (tng t nh mng trong c dng gp khc ty, lp th t bo nhn tht). CM c cha cc enzym ca chui h hp v cc enzym tng hp ATP (ATPsynthetase). cc vi khun lu hunh mu ta, CM cn cha b my quang hp. CM c mt cc enzym tham gia vo qu trnh tng hp lipid mng, peptidoglycan, acid teichoic, LPS, v cc polysaccharid n gin. Mt khc mng sinh cht cn l ni tng hp cc enzym ngoi bo, nm gia mng sinh cht v thnh t bo gip thu phn c cc cht dinh dng di dng polyme. Mng sinh cht tham gia vo qu trnh phn bo nh cc Mesosom. Mesosom l phn CM cun nhng vo trong t bo cht. Mesosom c loi nm gn mng sinh cht v loi nhng su vo t bo cht. Loi nm gn mng sinh cht c vai tr nht nh trong vic sinh tng hp ra penicilinase v mt s enzym khc. Cn loi Mesosom nhng su vo t bo cht c gn vi nhim sc th (NST) h tr qu trnh sao chp ADN v qu trnh phn bo. Ngoi ra mng sinh cht cn cung cp nng lng cho vn ng ca tin mao (lng roi, flagella). 1.2.3. T bo cht (cytoplasm) T bo cht (TBC) ca vi khun cha ti 80% nc di dng gel, cn li l cc cht ho tan nh protein, peptid, acid amin, hydratcarbon, lipid, cc nucleotid, ARN, vitamin, cc ion v c v nhiu cht c phn t lng thp khc. T bo cht ca vi khun khng di ng do c h thng cc mng li gip duy tr hnh dng ca t bo. c im ny khc hn vi TBC ca cc t bo nhn tht. Protein v polypeptid chim ti 50% khi lng kh ca vi khun v mt phn rt ln nng lng ca vi khun c s dng tng hp protein, trong cc protein cu trc v enzym ni bo c tng hp c hiu. Ribosom nm t do trong t bo cht, chim ti 70% khi lng kh ca cht nguyn sinh t bo vi khun. C ti 15.000 20.000 ribosom trong mt t bo E. coli. Ribosom bao gm 2 tiu phn 50S v 30S kt hp vi nhau to thnh monosom 70S (S l n v Svedberg, i lng o tc lng ng ca ht trong mt huyn dch khi ly tm siu tc). Mi tiu phn li bao gm 2 thnh phn polyme: protein v ARN l protein v ARN ribosom. Tiu phn ribosom 30S bao gm mt phn t ARN 16S v 21 phn t protein. Cn tiu phn 50S c cha 2 phn t ARN 16S nh tiu phn 30S v 34 phn t protein. Ngoi ra mt phn t ARN 5S cng c xc nh trong ribosom. Khi tng hp protein, cc ribosom gn vi mARN v c gi l polyribosom. Hnh 1.8 gii thiu s cu to
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ca ribosom. Ribosom ca vi khun chu tc dng ca nhiu loi khng sinh nh aminosid, tetracyclin, chloramphenicol,... Ngoi cc thnh phn nu trn, t bo cht cn cha cc th n nhp (cn gi l cc ht vi). y l cc khng bo cha lipid, glucogen v mt s khng bo cha cc cht c trng i vi tng loi vi khun. Chng hn vi khun bch hu c ht vi cha polymetaphosphat. So snh vi t bo cht ca sinh vt nhn tht, t bo cht ca vi khun khng c ty th, lp th, li ni bo v c quan phn bo. 1.2.4. Th nhn (Nuclear body) Vi khun l sinh vt procaryota, vi cu to nhn n gin, cha c mng nhn. Th nhn ca vi khun l mt nhim sc th (NST) c nht c cu to bi mt phn t ADN xon kp gn vi Mesosom. Phn ln cc VSV tin nhn l cc t bo n bi. Chiu di NST vi khun thay i trong khong 0,25 3,0m v nu dui ra thng di gp hng nghn ln chiu di t bo vi khun (chiu di ADN ca E. coli khong 1m). NST ca vi khun cha khong 6,6 13,0 106 cp base nit. NST ca vi khun E. coli c 4,6 106 cp base nit, khp kn, vi PTL l 3 109 Da, cha khong 3.000 gen.
Nhim sc th t bo tin nhn c hnh cu, hnh siu xon hay hnh que, sao chp theo m hnh bn bo tn (hnh 1.9). Tuy nhin s nhn ln ca vi khun cn ph thuc vo s phn chia ca
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mng t bo cht v thnh t bo, nhng NST bao gi cng c nhn ln trc. S lng h gen hay genom trong t bo vi khun thay i tu thuc vo loi v iu kin nui cy. Khi nui cy tnh t bo E. coli cha khong 1 4 genom, t bo Desulfovibrio gigas cha khong 10 15 genom. Ngoi nhim sc th mt s vi khun cn cha ADN ngoi nhim sc th. c th l cc loi plasmid v transposon. 1.2.5. V nhy (capsul) mt s loi vi khun bn ngoi thnh t bo cn c mt lp nhy lng, sn st, khng r rt bao quanh hay cn gi l capsul. Khi quan st di knh hin vi c th s dng phng php nhum mc tu s thy r v nhy mu trng ni ln trn nn sm. Khun lc ca nhng vi khun c v thng nhy, t v sng. Thnh phn ch yu ca v nhy l polysaccharid, ngoi ra cn c polypeptid v protein. V nhy polysaccharid chim t l ln (Streptococcus mutans, Streptococcus salivarius, Xanthomonas, Corynebacterium. V nhy ca Leuconostoc mesenteroides c cu to bi polysaccharid l dextran, polyme c sn xut tm thay th huyt tng cho ngi m mt nhiu mu. Sn phm dextran Fe c dng lm thuc b mu trong chn nui. V nhy ca mt s loi Bacillus (nh B. anthracis, B. subtilis,...) c cu to bi polypeptid, ch yu l acid polyglutamic. Cc chc nng ch yu ca v nhy l bo v, d tr tch lu v tng cng kh nng bm. Bo v vi khun trnh b thng tn khi kh hanh, trnh c hin tng thc bo (phagolysis) ca bch cu. V d, cc ph cu dng R (khng v) khng gy bnh v b thc bo, ngc li ph cu dng S nh c v bo v, lm bo ho opsonin ho, nn ngn cn thc bo m gy bnh. V nhy nh l kho d tr ngoi bo, khi thiu cht dinh dng, vi khun s dng v nhy nh ngun cht dinh dng. Mt khc v nhy cn l ni tch lu mt s sn phm trao i cht xung quanh t bo. Tng cng kh nng bm: Nh v nhy v mt s cu to b tr vi khun bm vo c b mt ca mt s gi th. V d, cc trng hp vi khun gy su rng Streptococcus salivarius v Streptococcus mutans sinh tng hp enzym hexoz transferase to ra polyme fructan t saccarose, gip vi khun bm c ln b mt ca rng, ln men ng sinh acid lactic v dn dn lm hng men rng, gy su rng. 1.2.6. Cc b phn ph khc Lng (roi, tin mao flagella): Lng l nhng si protein di un cong hoc xon c to thnh t cc acid amin dng D, c th quan st c di knh hin vi in t. Tin mao gip t bo chuyn ng ch ng c. Lng c ng knh khong 13,5 17,0nm v chiu di khong 10 20 m. Lng thng gp trc khun v xon khun (Vibrio, Spirilum, Pseudomoanas, Escherihcia coli, Sigella, Salmonella, Proteus,... cn Clostridium, Bacillus, Bacterium,... t gp hn), t khi gp cu khun (ch c chi Planococcus). Cu to ca lng c 3 phn: si, mc v gc. Si c cu thnh t mt s chui protein xon vi nhau to thnh v rng. Mc gi gn vo u cui ca si. Gc gn vo mc gi lng vo vch v mng t bo cht ca t bo. Tin mao to cho vi khun kh nng chuyn ng vi vn tc kh cao n khong 20m/s trong dung dch nc. Cc chuyn ng c th l dn d, xua ui, nho ln, tin,... Nhng vi khun c lng th c khng nguyn H (hnh 1.10)
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Khun mao (pilus, fimbria), cn c gi l nhung mao, l nhng si bn cht protein, rng, rt nh (ng knh trong 2,0 2,5nm, ng knh ngoi 7 9nm) v rt ngn, s lng c th rt nhiu (250 300 si/t bo), hay gp vi khun G. Cu to ca pili n gin hn ca lng.
Da vo chc nng c th chia khun mao lm hai loi: pili chung v pili gii tnh F. Pili (khun mao) chung l c quan bm, gip vi khun bm vo gi th ca cc t bo nhn tht (mng nhy ca ng h hp, ng tiu ho, ng tit niu, ng sinh dc ca ngi v ng vt,...) tng cng kh nng gy bnh ca VSV. Chng hn mt pili, lu cu khng cn kh nng gy bnh. Pili (khun mao) gii tnh (pili F fertility): Pili gii tnh ging nh pili thng nhng di hn nhiu. Ch c vi khun c c pili gii tnh v mi t bo c th c 1 4 pili gii tnh vi chc nng lm cu ni gia hai t bo chuyn ADN t t bo ny sang t bo khc khi tip hp. Ngoi ra pili gii tnh cn l th th cho cc Bacteriophage hp th (hnh1.11).
Bo t (nha bo spore endospore): Mt s vi khun vo cui thi k sinh trng khi iu kin pht trin khng thun li s sinh ra bn trong t bo mt th ngh c dng hnh cu hay hnh elipsoic, c gi l bo t (nha bo). Khi iu kin thun li, bo t li ny mm pht trin thnh t bo sinh dng. Bo t c hnh thnh trong cc bc sau: + Trong t bo sinh dng, ADN c phn chia thnh chromosom ring bit v mng t bo cht ln su vo phn chia t bo. + ADN c bao bc hon ton bng mng t bo cht, thnh nha bo c hnh thnh v mt phn v cng c hon thnh. + V bo t c hnh thnh y bao quanh ADN. + Bo t chn v gii phng ra ngoi t bo. Qu trnh hnh thnh bo t nu mt khong 6 8 ting, l qu trnh tng i phc tp
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(sporogenesis) to ra c cu trc vng bn, chu c cc iu kin khc nghit. Cu trc ca bo t l h thng nhiu lp v bn vng bao quanh vng li cha ADN. ADN nm trong th bo t cht trong cng, mng bo t bao ngoi bo t cht, tip n l thnh bo t, tip theo l v bo t, ri n o bo t hai lp v o ngoi. Cc lp bao bc chim 50% th tch ca nha bo v bao bc bo t cht v nhn. Thnh phn ho hc ca cc lp v bao l protein, hydratcarbon, lipid, peptidoglycan v calci dipicolinat (CaDPA), khng cha acid teichoic. Li ca bo t cha mt lng nc rt thp cng vi CaDPA. CaDPA c hnh thnh trong qu trnh nha bo ho v mt i khi bo t ny mm. Trong bo t cn cha nhiu enzym chuyn ho nhng dng khng hot ng v chng ch bt u hot ng khi bo t ny mm. Bo t c tnh khng nhit, khng bc x, khng ho cht, khng p sut thm thu. Chng hn, bo t ca vi khun gy ng c tht Clostridium botulinum b em un si 100OC trong 5,0 9,5 gi mi cht, hp 121OC sau 10 pht mi b cht. Bo t c th gi sc sng t vi nm n vi chc nm. c nhng thng bo tm thy c nhng bo t trong cc tiu bn kho c cch y 3.000 nm m vn duy tr c sc sng. Ch mt s chi vi khun c kh nng sinh nha bo: Bacillus, Clostridium, Sporosarcina (G+) v Desulfotomaculum (G). Bacillus v Clostridium l trc khun, Sporosarcina l hnh khi 8 cu khun, v Desulfotomaculum (G) l vi khun kh lu hunh, t bo hnh que cong, ring r hay lin kt thnh chui. Hnh 1.12 gii thiu bo t vi khun.
Khi gp iu kin thun li v c hot ho bi mt s yu t lm tn thng cc lp bao nh nhit , ho cht (acid, cc cht cha nhm SH t do) th v ca bo t b (t) phn hu (autolysis), bo t hp th nc phng ln, gii phng CaDPA, cc enzym phc hi hot ng, h hp v sinh tng hp cc cht tng nhanh, sau khong 4 5 gi, bo t tr thnh t bo sinh dng. 1.2.7. Phn loi vi khun Phn loi VSV gip h thng ho c cc VSV ngy cng phn lp c nhiu hn, a dng hn. Tuy nhin, phn loi cc VSV ng tin cy, chnh xc, i hi nhiu cng sc. 1.2.7.1. Cc phn nhm phn loi Cc phn nhm phn loi ca VSV nm trong h thng phn loi ca th gii sinh vt ni chung, bao gm: 1. Gii (kingdom): V d: Protista. 2. Ngnh (division hoc phylum), di ngnh (subdivision). 3. Lp (class): V d: Actinomycetes, di lp (subclass). 4. B (order): Gi tn khoa hc ly tn h chnh thm v t ales. V d: Pseudomonadales.
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B ph (suborder) hay di b, tn cng bng ineae. 5. H (family): Gi tn khoa hc vi v t aceae, v d: Enterobacteriaceae. Di h: gi tn khoa hc tn cng bng oideae. 6. Tc (tribe): gi tn khoa hc thm v t eae, v d: Escherichieae. Di tc (subtribe), gi tn khoa hc thm v t inea. 7. Chi (Genus): V d: Proteus, Staphylococcus,... 8. Loi (species): y l n v phn loi c bn nht. Tn khoa hc ca cc loi VSV thng gi kp, tn chi trc v tn loi sau: v d: Staphylococcus aureus. 9. Th (variety): Ch mt nhm nht nh trong loi, v d Mycobacterium uberculosis var. hominis trc khun lao ngi. 10. Dng (typ hoc forma): Ch nhm nh di th, v d: Streptococcus pneumoniae typ 14. 11. Chng (strain): Ch mt tp on VSV ng nht v hnh thi v sinh l,... ca mt loi mi c phn lp, mang theo k hiu ca chi, loi v chng. V d: Staphylococcus aureus ATTC 1529. Trong VSV hc ch yu s dng cc phn nhm phn loi lp, b, h, chi, loi, th, typ v chng. 1.2.7.2. Cc phng php phn loi Phn loi vi khun c th tin hnh theo mt s phng php sau: 1. Phn loi theo s lng cc tnh cht sinh hc (phng php kinh in): Xc nh cc c trng phn loi, nu ging nhau ln hn 90% th cng nm trong mt loi. 2. Phn loi theo phng php phn t: So snh thng tin di truyn trong ADN ca cc VSV khc nhau, bao gm: + Theo t l cc base ca ADN. V d: (G + C)/(G + C + A + T). + Lai ADN (hoc lai cADN t mARN). + Xc nh trnh t ADN. + Lai sinh hc. + Phn loi da trn cu trc protein.
2. X KHUN (ACTINOMYCETES)
Trc th k XIX, x khun c xp vo nhm nm, tuy nhin, khi nghin cu su hn xc nh c cc x khun c nhn nguyn thu v kch thc nh, do chng c xp vo nhm vi khun tht.
X khun thuc nhm vi khun tht phn b rng ri trong t nhin, l cc vi khun G+, c t l G + C > 55%. Trong mi gam t ni chung thng c cha hng triu x khun. i a s x khun l cc VSV hiu kh, hoi sinh, c cu to dng si phn nhnh. Do c th sinh tng hp c nhiu sn phm trao i cht quan trng, nn cc x khun c cc nh khoa hc quan tm nghin cu rt nhiu. Trong s hn 10.000 khng sinh hin c bit trn th gii th khong 66% l do x khun to ra. X khun cn c s dng iu ch nhiu loi enzym (amylase, cellulase, glucoizomerase, protease,...), cng nh cc hp cht khc. X khun gy bnh cho ngi hoc ng vt l trng hp rt hn hu. Hnh 1.13 gii thiu hnh dng cc chui bo t ca x khun.
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Hnh 1.13. Hnh dng cc chui bo t ca x khun
2.1. Cc c im hnh thi ca x khun

Nh nu trn, x khun thuc nhm cc vi khun tht, nhng pht trin thnh dng si. H si ca x khun chia ra thnh khun ty c cht v khun ty kh sinh. Khun ty c cht l khun ty c bn, cn khun ty kh sinh pht trin mnh hay yu, thm ch hu nh khng pht trin tu tng chi, tng loi. Tp hp mt khm x khun pht trin ring r to thnh khun lc x khun. Khun lc x khun kh c bit, khng trn t nh vi khun, nm men m thng c dng th rp, dng phn, khng trong sut, c cc np gp to ra theo hnh phng x (Actinomycetes ting Hy Lp: Aktis, aktinos l tia, mykes l nm), dng que cy khng di chuyn c khun lc x khun v khun ty c cht bm su vo trong thch. i vi nhng nh thng tho trong chuyn ngnh khun lc ca x khun khng th lm ln c vi cc nm lc ca vi nm v si kh sinh ca nm thng c ng knh ln hn khun ty kh sinh ca x khun 5 10 ln. ng knh khun ty x khun thay i trong khong t 0,3 1,0m n 2 3m. a s khun ty x khun khng c vch ngn. Mu sc khun ty x khun ht sc phong ph. C th gp cc mu da cam, en, , lc lam, nu, trng, vng, xm,... Khun ty c cht c th tit vo mi trng mt s loi sc t, c sc t tan trong nc, c loi ph thuc pH, c loi ch tan trong dung mi hu c. Trong mi trng c hiu, c loi x khun c th to ra sc t melanoid sm en. Khun ty c cht ca x khun pht trin mt thi gian th di ra trong khng kh thnh nhng khun ty kh sinh. Ngi ta cn gi khun ty kh sinh l khun ty th cp phn bit vi khun ty s cp l loi khun ty bt u pht trin t cc bo t ny mm. i vi cc x khun thuc h Streptomycetaceae, sau mt thi gian pht trin, trn nh khun ty kh sinh s xut hin cc chui bo t. Chui bo t c th mc n hay mc vng gm cc hnh thi c bn nh: thng, un cong, mc cu n hoc kp, v xon l xo. Bo t trn (conidiospore) ca h x khun ny c cc hnh dng: hnh cu, hnh elipsoid bu dc, hnh tr,... B mt bo t c th l nhn, sn si da cc, c gai hoc c tc. Bo t trn l c quan sinh sn ch yu ca h x khun ny. Bo t trn c hnh thnh theo hai phng thc khc nhau: Vch ngn hnh thnh t bn trong ca CM v tin dn vo trong to ra nhng vt ngn khng hon chnh sau chui bo t mi c phn ct thnh cc bo t trn. Thnh t bo v CM ng thi xut hin vch ngn tin dn vo pha trong v lm cho si tin chui bo t phn ct to thnh chui bo t trn.
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H Actinoplanaceae to ra cc bo t bn, c kh nng di ng, cn h Actinomycetaceae to ra cc bo t hnh cu bng cch phn ct h si trng thnh gi. Mt s x khun c sinh nang bo t bn trong c cha cc bo t nang.
2.2. c im cu to ca t bo
T bo x khun c mt s c im khc t bo vi khun: Thnh t bo x khun c dng kt cu li, dy khong 10 20nm, c chc nng duy tr hnh dng ca khun ty v bo v t bo. Cn c vo thnh phn ho hc, thnh t bo x khun c chia ra lm 4 nhm: + Nhm CW I: C cha LDAP (Ldiaminopimelic acid) v glycin, bao gm Nocardioides, Sporichthya, Streptomyces v Streptoverticillum. + Nhm CW II: C cha mesoDAP (mesodiaminopimelic acid) v glycin, bao gm Actinoplanes, Ampullariella, v Micromonospora. + Nhm CW III: Ch cha mesoDAP, bao gm Actinomandura, Actinosynnema, Dermatophyllus, Frankia, Geodermatophyllus, Microbispora, Nocardiopsis, Streptosporangium, Planobispora, Planomonospora, Thermoactinomyces, Thermomonospora. + Nhm CW IV: Cha mesoDAP, arabinose v galactose, bao gm Actinopolyspora, Mycobacterium, Nocardia, Oerskovia, Promicromonospora, Pseudonocardia, Rhodococcus, Saccharomonospora, Saccharopolyspora. Mng t bo cht ca x khun dy khong 7,5 10nm. Chng c cu trc v chc nng ging nh ca vi khun ni chung. Mesosom nm pha trong ca t bo cht, c hnh phin, hnh bng hay hnh ng. Mesosom lm tng din tip xc ca CM v qua lm tng cng hot tnh enzym, tng vn chuyn in t,... Cc vt th n nhp trong t bo cht ca x khun gm c cc ht phosphat (dng hnh cu, bt mu vi thuc nhum Soudan III), cc ht polysaccharid (bt mu vi dung dch Lugol).
2.3. Mt s kho phn loi v cc c im phn loi ca x khun

C kh nhiu kho phn loi x khun c cc nh khoa hc nghin cu pht trin phn loi cc x khun ngy cng nhiu thm, trong phi k n kho phn loi ca Waksman, kho phn loi ca Krassilnhikov (Nga), kho phn loi ca Gauze,... Cc kho phn loi chia lp x khun (Actinomycetes) thnh lp ph (hoc b) Actinomycetales v cc VSV ging x khun (like Organisms). Lp ph Actinomycetales c chia thnh cc h: Actinoplanaceae, Actinomycetaceae, Streptomycetaceae, Micromonosporaceae, Nocardiaceae,... Kho phn loi Streptomyces thuc Chng trnh Streptomyces quc t (International Streptomyces Project) do Shirling v Gotlieb xut (1970) c s dng rng ri phn loi cc x khun thuc chi Streptomyces trong h Streptomycetaceae. Trong kho phn loi ny, cc c trng phn loi c s dng bao gm: mu sc khun lc, mu khun ty c cht, sc t melanoid, sc t ho tan, hnh dng chui bo t, b mt bo t (chp nh knh hin vi in t), tiu th ng arabinose, tiu th ng xylose, tiu th ng inositol, tiu th ng manitol, tiu th ng fructose, tiu th ng rhamnose, tiu th ng saccarose, tiu th ng raffinose. Chng hn, cc c trng phn loi ca Streptomyces rimosus sinh tng hp oxytetracyclin c c trng ln lt (theo th t c nu trn) nh sau: W Y(R) 0 1 0 S sm + + + + +. Hng nm, s tay ca Bergey v Vi khun hc (Bergey Manuals of Becteriology) vn cng b phn loi cc x khun theo cc c trng phn loi ch yu gn ging nh nu trn, l ch dn quan trng m cc nh phn loi x khun quc t c th tham kho.
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Tuy nhin ngy nay, vic p dng k thut xc nh trnh t gen phn loi x khun cng v ang c p dng, nht l cc phng th nghim tin tin. u im l cho kt qu kh chnh xc, ng tin cy. Yu im l lm cho cc nh nghin cu tr nn giy t, xa l hay h hng vi cc c trng sng (m y cng chnh l biu hin ca cc sn phm gen) ca VSV.
3. NHM CC VI KHUN NGUYN THU

Nhm vi khun ny l nhm vi khun c kch thc nh gm 3 loi gm Chlamydia, Mycoplasma v Rickettsia. Cc vi khun ny c th chim v tr trung gian gia virus v vi khun. Bng 1.2 gii thiu mt s c im ca Chlamydia, Mycoplasma v Rickettsia, so snh vi virus v vi khun.
3.1. Cc Chlamydia
Nm 1907, cc nh khoa hc ln u tin pht hin ra th bao hm trong t bo kt mc ca bnh nhn au mt ht (trachoma). Thut ng Chlamydia (t gc Hy Lp Chlamys l o chong) c Jones, Rake v Steans xut nm 1945. Sau nhiu ln i tn, n nm 1970, ti Hi ngh quc t v mt ht ti M, nhm VSV ny chnh thc c gi tn l Chlamydia. Hi chng Chlamidiosis gm 3 bnh: st vtst chim (ornithosepsittacose), bnh NicolasFaviruse v bnh mt ht. Ngy nay, Chlamydia cc cn gy ra mt s bnh ng sinh dc tit niu ang c quan tm. Cc Chlamydia l loi vi khun rt b nh, qua lc, Gram(+), c chu k pht trin c bit, k sinh bt buc trong t bo ng vt nhn thc: chut trng, bo thai g, pht trin trn t bo c nui cy (t bo thn kh), trong trng g p, nht l mng niu m, ti non hong. Chlamydia khc virus cc im sau y: C cu to t bo, c thnh t bo cha peptidoglycan c trng cho vi khun G+. C ribosom trong t bo cht. Sinh sn bng phn i n gin. C cha ng thi hai loi acid nucleic ADN v ARN. H thng enzym trao i cht khng hon chnh, thiu cc enzym tham gia trao i nng lng, do bt buc phi k sinh trong cc t bo nhn thc. Rt mn cm vi cc cht khng sinh v cc sulfamid (ring Chlamydia psittaci c tnh khng cao i vi sulfamid). Trong phng th nghim c th nui Chlamydia trong mng bao lng trng g, trong khoang bng chut bch, trn t bo Hela,... Chlamydia c chu k sng c bit: Dng c th c kh nng xm nhim gi l nguyn th nguyn bo. l dng t bo hnh cu c th chuyn ng, ng knh rt nh (0,20,5m), nhum
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Giemsa bt mu tm, nhum Machiavello bt mu . Nguyn th xm nhp vo trong t bo, phn mng bao quanh nguyn th bin thnh khng bo. Nguyn th ln dn ln trong khng bo v bin thnh thu th. Thu th (thu = nguyn thu) cn gi l th dng li, l loi t bo hnh cu, mng mng, kh ln, ng knh (0,8 1,5m). Thu th lin tip phn chia thnh hai phn u nhau v to thnh vi khun lc trong t bo cht ca vt ch. Cc t bo con ny li phn ho thnh cc nguyn th nh hn na, mng dy v c tnh cm nhim. Khi t bo vt ch b ph v, cc nguyn th c gii phng ra s xm nhim vo cc t bo khc (hnh1.14).
Theo Bergey, h Chlamydiaceae c hai chi: Miyagawanella v Chlamydia. Nhiu loi Chlamydia gy bnh nguy him cho ngi v ng vt: Miyagawanella psittaci (Chlamydia psittaci) gy bnh st vt. Miyagawanella lymphogranulomatosis gy bnh vim hch bch huyt hoa liu bn. Chlamydia hoc Chlamydozoon oculogienitalis gy vim niu o, c t cung v kt mc th vi. Chlamydia trachomatis gy bnh mt ht ngi v chut nht. Loi ny c ba bin chng:
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BT (Biovar trachoma), LGV (Biovar lymphogalunoma venerum) v BM (Biovar mouse). Bin chng Chlamydia trachomatis BT gy bnh au mt ht cho ngi t cch y t ra l 3.000 nm. Ngy nay trn th gii c khong 400 triu ngi au mt ht, v c khong 6 10 triu ngi b m v bnh mt ht ny. Bin chng Chlamydia trachomatis LGV gy ung th ht ng sinh dc (ly truyn qua ng tnh dc).
3.2. Cc Mycoplasma
Mycoplasma u tin c pht hin b b vim phi mng phi vo nm 1898 nh Nocard v Roux, c t tn l Mycoplasma mycoides. Sau nhiu Mycoplasma gy bnh cho ng vt c phn lp. Nm 1937, Edsarr v Dienes phn lp c Mycoplasma hominis tuyn Bartholin ngi. Nm 1961, Marmior v Goodburd Chanock v Hayflick, 1962 chng minh c tc nhn gy bnh vim phi khng in hnh ngi l Mycoplasma ch khng phi l virus, v c t tn l Mycoplasma pneumoniae. Ngy nay nhiu Mycoplasma gy bnh sinh dc tit niu c phn lp (hnh 1.15).
Hnh 1.15. Mycoplasma hominis (ln, nht, dng trng p lp) v Ureaplasma urealiticum (nh mu m)
c im sinh hc
Mycoplasma l nhng vi khun rt nh, nh nht trong cc VSV sng c lp, khng di ng, khng sinh bo t, hnh thi rt a dng (hnh thoi, hnh que ngn hoc hnh cu). Hnh th ca Mycoplasma thay i ph thuc vo tui ca qun th VSV v yu t mi trng. Mycoplasma khng bt mu Gram, rt kh nhum, quan st di knh hin vi cho kt qu khng chc chn. Mycoplasma c th pht trin trn cc mi trng c hay khng c t bo sng. Trong mi trng khng c t bo sng, cn b sung nhng cht dinh dng c bit nh huyt thanh nga, cao nm men. Mycoplasma c nhiu loi k kh hay hiu kh bt buc nhng cng c loi k kh tu tin. Nhit ti u l 35 37OC, pH l 7,0 7,8. Trong mi trng lng, khng lm c mi trng nn kh nhn bit. Trn mi trng c, to khun lc b, phi s dng knh lp hay knh hin vi quan st: mng v dt nh trng p lp, hnh dng in hnh khi t thch. V ngoi ca Mycoplasma yu, ch nh mng t bo cht ca cc VSV khc, tuy nhin rt do, v th nn hnh dng t bo hay thay i. Di knh hin vi in t, mng t bo c cu to dng ht hoc dng li vi cc ribosom. Qu trnh nhn ln ca Mycoplasma rt phc tp: phn i kt hp vi ny chi, nu trong t bo nui th pht trin trn b mt t bo. Mycoplasma d b tiu dit bi nhit, ho cht, siu m, tuy nhin khng c penicilin. Mycoplasma c cc yu t khng nguyn l lipid hay polysaccharid. Theo h thng phn loi Bergey, b Mycoplasmatales c 3 h. H Mycoplasmataceae bao gm chi
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Mycoplasma v chi Ureaplasma. Cc Mycoplasma gy bnh cho ngi bao gm: M. fermentans, M. hominis, M. pneumoniae, M. orale hoc M. pharingis, M. salivarium, M. urealiticum. Mycoplasma pneumoniae gy vim phi tin pht khng in hnh, Mycoplasma orale gy vim ng sinh dc tit niu n ng, Mycoplasma urealyticum gy vim niu o khng do lu v Mycoplasma hominis gy nhim trng ph khoa. C th s dng cc khng sinh tetracyclin, chloramphenicol, spiramycin, cng nh doxycyclin, cefalexin, cefotaxim iu tr bnh do Mycoplasma c hiu qu tt.
3.3. Cc Rickettsia
Nm 1909, H.T. Ricketts (1871 1910) ln u tin pht hin ra mm bnh ca bnh st thng hn pht ban (sau ny gi l bnh st Ricketxi pht ban). Nm 1910, ng hy sinh trong khi nghin cu bnh ny. Tn Rickettsia c t cho nhm VSV ny l ghi nh cng lao ca ng. Rickettsia l VSV nhn nguyn thu, Gram(), ch c th tn ti trong t bo cc VSV nhn tht. Rickettsia khng th sng c lp trong cc mi trng nhn to, khng c dng qua lc, nng lc sinh tng hp kh mnh v khng to thnh cc th bao hm. n nay c vi chc loi Rickettsia c pht hin. Rickettsia c cc c im chung sau y (hnh 1.16):
Hnh 1.16. Rickettsia rickettsii trong t bo vt ch
Rickettsia l vi khun khng di ng, t bo c kch thc khong 0,5 1,0m. Hnh thi t bo Rickettsia lun bin i, nhng hay gp nht l dng trc khun, nhng cng c th l hnh cu, hoc que ngn,... Trong t bo b xm nhim, Rickettsia sp xp v quy tc nhng thng t tp thnh tng khi dy c. Rickettsia l vi khun Gram(), kh nhum, khi nhum Giemsa bt mu hng, nhum Macchiavello bt mu fuchsin trn nn xanh (tr R. tsutshugamushi bt mu xanh). Rickettsia k sinh c ng vt v thc vt. Rickettsia k sinh bt buc trong t bo cc sinh vt nhn thc. Vt ch thng l cc ng vt c chn t nh ve, bt, b, rn,... Rickettsia sinh sn bng phng thc phn i n gin. Rickettsia ni chung mn cm vi nhit , 56OC sau 30 pht thng b cht. Chu trnh trao i nng lng ca Rickettsia khng hon chnh, phn ln ch c th s dng acid glutamic sinh nng lng ch khng th s dng c ng glucose. V mc ch nghin cu, cc Rickettsia thng c nui cy trong phi g, trong cc ng vt mn cm, trong cc t chc nui cy (nh dng t bo Hela). Theo h thng phn loi Bergey, tt c Rickettsia c phn vo b Rickettsiales. Trong b ny
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c tt c 3 h v 14 chi, ng ch hn c l h Rickettsiaceae, trong c cc loi Rickettsia gy bnh ch yu ngi gm: St pht ban dch t: Rickettsia prowaseki gy bnh st Rickettxi pht ban; R. mooseri (hay cn gi l R. typhi) gy bnh Rickettxi st pht ban chut, hay st pht ban a phng. St do ve truyn: Dermacentrocenus rickettsia gy st pht ban v st c tnh Braxin v Nam Phi; D. conori, D. rickettsia gy st a Trung Hi; D. rickettsia gy st ng Phi v Nam Phi; D. rickettsia gy st ve truyn Bc ; bin chng D. rickettsia gy st ve truyn Bc c. St m truyn: R. tsutsugamushi (hay cn gi l Rickettsia orientalis) gy bnh tsutsugamushi. Gy bnh st hm ho (trench fever): Rochalimaea quyntana gy bnh st chin ho, hay cn gi l st nm ngy. Gy bnh st Q: Coxiella burnetii gy bnh st Q, hay cn gi l st Query; Rickettsia hay Dermacentrocenus gy bnh st cc do ve truyn. Rickettsia rickettsii gy bnh st pht ban ni Rocky Mountain spotted fever. Gy bnh cho ng vt c R. ruminantium gy trn dch mng tim ca cc ng vt c sng. Nhim khun Rickettsia c kh nng min dch mnh m v lu di, nu mt ln mc bnh, ln tip xc sau khng b mc li bnh. Mc bnh do Rickettsia gy ra thng c biu hin st, km theo pht ban in hnh da, c bit trong nhiu trng hp u c tn thng mch mu nh kiu vim hoc vim tc mao mch. Cc Rickettsia mn cm vi cc khng sinh nh lincomycin, fluoroquynolon nn c dng iu tr bn cnh tetracyclin, cloramphenicol. i vi tr em v ph n mang thai s dng rovamycin.
4. VI KHUN LAM
Vi khun lam (Cyanobacteria) trc y thng c gi l to lam (Cyanophyta hay blue algae) hay to lam lc (blue green algae), nhng thc t y l nhm vi khun c kh nng t dng quang nng, nh cha dip lc a. Qu trnh quang hp ca vi khun lam l qu trnh phosphoryl ho quang hp phi tun hon, c gii phng oxy nh cy xanh, khc vi qu trnh phosphoryl ho quang hp tun hon khng gii phng oxy nh vi khun k kh mu ta khng cha lu hunh thuc b Rhodospirillales. Vi khun lam phn b rng ri trong t nhin, ch yu sng trong nc ngt v to thnh thc vt ph du ca cc thu vc. Mt s phn b trong nc mn v nc l giu cht hu c, mt s sng cng sinh, v d Anabaena azollae cng sinh trong bo hoa du,... Nhiu vi khun lam c kh nng c nh nit (Nostoc, Anabaena, Gloeotrichina, Tolypothrix, Scytonema,...) c th cy nhim vo cc rung la cho cht lng tt. Sinh khi mt s vi khun lam c gi tr dinh dng cao (Spirulina cn gi l to Spirulina) nn c sn xut vi quy m cng nghip, gp phn ci to mi trng v ngun thc n chn nui. Theo h thng phn loi ca Bergey, vi khun lam c chia lm 5 b: b Chrococcales c cc chi Chamaesiphon, Gloeobacter, Gloeothece, n bo hay kt thnh tp on, sinh sn bng phn i. B Pleurocapsales c cc chi Dermocarpa, Xenococcus, Dermocarpella, Myxosarcina, Chroococcidiopsi: n bo phn chia nhiu ln, c th to si hay tn (thallus). B Oscillatoriales gm cc chi Spirulina, Arhtrospira, Oscillatoria, Lyngbya, Pseudanabaena, Starria, Crinalium, Microcoleus: a bo dng si, khng c t bo d hnh. B Nostocales gm cc chi Anabaena, Aphanizomenon, Nodularia, Cylindrospermum, Nostoc, Scytonema, Calothrix: a bo dng si, c cc t bo d hnh tham gia c nh nit. B Stigonematales c cc chi Chlorogloeopsis, Fischerella, Stifonema, Geitleria: a bo dng si, thng c dng tn (heterotrichous), c s phn chia ngang v thng. Vi khun lam khc bit rt ln vi to: khng c lc lp, khng cha nhn thc, c ribosom 70S, thnh t bo c cha peptidoglycan, nn mn cm vi penicilin v lyzozym. T bo sinh dng c hnh cu, hnh elipsoid,... ng knh c khi ch 1m (Synechococcus), nhng c khi n 30m (Oscillatoria). Cu to t bo vi khun lam gn vi vi khun Gram (). C quan thc hin qu trnh quang hp l tilacoit (thylacoids), dng bn xp song song hay un cong, c cha clorophil a, caroten,... Hnh thi ca vi khun lam c nhng dng kh c bit nh t bo d hnh (heterocyst), bo t ngh (akinete), to on (hormogonia), vi tiu bo nang (mannocyst), ht sinh sn (gonidium),... cc vi khun lam cha pht hin thy hnh thc sinh sn hu tnh.
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CC CU HI LNG GI
1. 2. 3. 4. 5. Trnh by cc c im hnh thi ca cc loi vi khun chnh. V v gii thch s cu to ca t bo vi khun Procaryota. Trnh by cc thnh phn, cu to v chc nng thnh t bo cc vi khun G+ v G. Trnh by chc nng, thnh phn v cu to ca mng t bo cht, th nhn v bo t ca VSV Procaryota. Trnh by cc c im chnh ca cc x khun, cc vi khun nguyn thu, vi khun lam.
Chng 2
DINH DNG VI SINH VT
1. CC CHT DINH DNG CA VI SINH VT

D nh b nhng cc VSV l nhng c th sng c lp i hi a dng cc cht dinh dng cho hot ng sng ca mnh. Cht dinh dng l nhng cht c VSV hp th, c tham gia vo cc qu trnh trao i cht ni bo ca VSV. Nhu cu dinh dng ca VSV l rt ln. Nu nh con ngi cn lng thc n khong 1/10 so vi khi lng c th, th VSV cn lng cht dinh dng bng chnh khi lng ca chng. Nhng nhm cht dinh dng m VSV cn thit bao gm ngun carbon, ngun nit, cht khong v cc cht sinh trng.
1.1. Ngun dinh dng carbon ca vi sinh vt

Cn c vo ngun dinh dng carbon, ngi ta chia VSV vo cc nhm sinh l gm cc VSV t dng carbon v cc VSV d dng carbon.
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1.1.1. Cc VSV t dng carbon Cc VSV t dng carbon s dng c ngun carbon v c, c th l CO2. T dng quang nng: Ngun carbon l CO2, ngun nng lng l nh sng: sinh oxy l nhm vi khun lam, khng sinh oxy l cc vi khun thuc chi Chromatium, Rhodopseudomonas, Chlorobium, hay Chloroflexus,.... T dng ho nng: Ngun carbon l CO2, ngun nng lng l mt s hp cht v c n gin. a s l cc vi khun hiu kh, tuy nhin, mt s vi khun k kh s dng nitrat hoc carbonat. Mt s vi khun lu hunh nh Thiobacillus thioparus, T. denitrificans, T. ferrooxidans cn c th phosphoryl ho c cht sinh nng lng. 1.1.2. Cc VSV d dng carbon Cc VSV d dng carbon khng s dng c ngun carbon v c, m phi s dng ngun carbon hu c. D dng quang nng: Ngun carbon l cht hu c,... ngun nng lng l nh sng, v d nh vi khun khng lu hunh mu ta. D dng ho nng: Ngun carbon l cht hu c, ngun nng lng thu c t chuyn ho trao i cht nguyn sinh ca mt s c th khc. V d: nm v mt s vi khun. Hoi sinh: Ngun carbon l cht hu c, ngun nng lng thu c t s trao i cht nguyn sinh ca cc xc hu c, v d nhiu nm v vi khun. K sinh: Ngun carbon l cht hu c, ngun nng lng ly t cc t chc hoc dch th ca mt c th sng. Cc VSV gy bnh cho ngi, ng vt, thc vt. Nh vy, tu nhm VSV m ngun carbon cn c cung cp c th l cht v c (CO2, NaHCO3, CaCO3,...) hoc cht hu c. Gi tr dinh dng ca cht dinh dng ch c ngha khi ph hp vi kh nng hp th cc ngun dinh dng carbon khc nhau ca nhm i tng VSV c cung cp. iu ny ph thuc vo thnh phn ho hc v tnh cht sinh l ca cht dinh dng, cng nh c im sinh l ca loi VSV quan tm. Theo thi gian, trn th gii, tt c cc hp cht carbon hu c u s b hoc nhm VSV ny hoc nhm VSV khc phn gii. Nhiu VSV c th ng ho c c cc hp cht carbon rt bn vng nh cao su, cht do, du m, parafin, kh thin nhin. Ngay formaldehyd l ho cht dit khun, nhng cng c nhm nm si s dng lm ngun dinh dng c. Nhiu cht polyme hu c c phn t lng qu ln VSV khng hp th c, nn VSV phi tit ra cc enzym (amylase, cellulase, pectinase, lipase,...) thu phn ngoi bo phn gii chng thnh cc hp cht phn t lng nh d hp th (ng n, ng i, acid bo,...). Cc loi sn phm c hm lng tinh bt cao nh bt ng, bt go, tinh bt cc loi c th s dng lm ngun carbon cho VSV. Tuy nhin, khi s dng cc loi bt cha tinh ch hm lng chnh xc ca mi trng khng xc nh c. Cc loi ng (ng n, ng i) thng c cc VSV s dng lm ngun dinh dng carbon sau khi c gii phng t thu phn cc polysaccharid. Trong phng th nghim, vic s dng cc ng tinh khit lm ngun carbon cho cc VSV l rt thng dng, v cho bit r c hm lng ca thnh phn mi trng, tuy nhin, i khi gi cc loi ng khng phi l r. Ngun hydratcarbon c kh nhiu nm men, nm mc v x khun a s dng l dch chit malt c cha ch yu l maltose v glucose v mt s protein, peptid v acid amin, trong prolin chim khong 50%. Trong cng nghip, r ng (d phm khng kt tinh c ca cc nh my ng) l ngun nguyn liu carbon r tin v rt thch hp vi nhiu loi VSV. Tuy nhin, r ng l ngun nguyn liu khng chun ca cng nghip khng sinh. Phm vi ng ho cc cht dinh dng carbon ca tng loi VSV rt khc nhau. C loi VSV ng ho c rt nhiu loi cht dinh dng nh Pseudomonas cepacia c th ng ho c 90 loi cht dinh dng carbon, trong khi c loi VSV sinh methan ch ng ho c CO2, hoc cc cht c cha 1 carbon hoc 2 carbon m thi. Vi VSV d dng, ngun dinh dng carbon c s dng vo c hai mc ch: ngun dinh dng v ngun nng lng.
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Mt s VSV k sinh, nht l cc vi khun gy bnh trong mu, trong cc t chc hoc trong rut ca ngi v ng vt, mun sinh trng c, ngoi ngun carbon hu c cn cn phi c cung cp mt lng nh CO2.
1.2. Ngun dinh dng nit ca VSV

+ Ngun nit d hp th nht i vi VSV l NH3 v NH 4 . Mui amoni ca cc acid hu c cng
thng c s dng. Tuy nhin, trong cng nghip sn xut cc sn phm th cp, khi s dng cc mui amoni hp th nhanh phi ch c ch trao i cht lm gim thiu, thm ch nh ch sn xut hot cht mc tiu. Ure l ngun dinh dng nit trung tnh v mt sinh l, khi b phn gii bi enzym urease s gii phng nhanh NH3 v CO2. Phn NH3 s c VSV s dng tip. H2NCONH2 + H2O 2NH3 + CO2 Mui nitrat l ngun dinh dng nit thch hp cho nhiu loi nm si, x khun v to, tuy nhin, khng thch hp lm cho nm men v vi khun. Sau khi VSV s dng ht gc NO3 , cc ion kim loi cn li s lm mi trng b kim ln. trnh hin tng ny, ngi ta s dng NH4NO3
NO3 .
+ lm ngun nit cho nhiu loi VSV. Tuy nhin, NH 4 thng c hp th trc ri mi n
Ngun nit d tr to ln trong t nhin l kh nit t do trong kh quyn, nhng c lin kt ba (N N) rt bn vng nn a s VSV khng c kh nng ng ho nit trong khng kh. Tuy nhin, c nhng VSV c th c kh nng chuyn ho N2 thnh NH3 nh hot ng xc tc ca h thng enzym nitrogenase. Cc VSV c kh nng ny gi l cc VSV c nh nit (nitrogen fixing microorganisms). Cc VSV thuc chi Rhizobium, chi Bradyrhuzobium c kh nng to nt sn trn cc cy h u nh R. leguminosarum ( u H Lan), R. phaseoli ( u xanh, u ty), B. japonicum ( u tng). Mt s x khun thuc chi Frankia c kh nng c nh nit trong cc cy khng phi h u. Xon khun Azospirillum lipoferum, A. brasilence,... sng trong r mt s c nhit i cng c kh nng c nh nit tt. Cc Azotobacter nh Azotobacter chrococcum, A. vinelandii c kh nng pht trin v c nh nit tt khi pH, hm lng calci v phospho thch hp. Ging vi cc Azotobacter l cc Beijerinckia nh Beijerinckia india, Beijerinckia fluminensis c th pht trin c nh nit pH mi trng kh thp, thm ch n 3,0. Vi khun k kh t do Clostridium pasteurianum, C. acetobutilinum, C. multifermentans,... cng c kh nng c nh nit phn t. Mt s vi khun lam nh Anabaena azollae cng sinh trong bo hoa du cng c kh nng c nh nit. Li dng kh nng c nh nit ca cc VSV trn nhm tng cng sn lng cy trng l n lc hu ch trong nng nghip. Nhiu VSV c kh nng ng ho tt nit cha trong cc hp cht hu c. Cc cht ny va l ngun nit, va l ngun carbon cho VSV. Nhng VSV khng c kh nng hp th trc tip cc protein cao phn t, m ch c cc olygopeptid cha khng qu 5 gc acid amin mi c th c hp th trc tip qua mng t bo cht ca VSV. B li, rt nhiu VSV c kh nng to ra protease cng nh peptidase ngoi bo, xc tc thu phn protein thnh cc hp cht peptid phn t nh v acid amin, c kh nng hp th c vo t bo VSV. Ngun nit hu c thng c s dng nui cy VSV l bt u tng, cao ng, cao nm men, cao tht, caseinhyrolysat v pepton,.... Cc ngun dinh dng hu c c cha c carbon v nit (pepton, nc tht,...) c th s dng va lm ngun carbon va lm ngun nit i vi VSV. Pepton l ch phm thu phn cha trit ca ngun protein no . Tuy nhin, trong cng nghip v l do kinh t k thut m trong s cc ngun nit hu c rt tt nu thng ch bt u tng v cao ng l c ng dng; cng nh thay cho cao nm men rt t nm men ti li c a dng. Xt v nhu cu acid amin c VSV t dng acid amin, VSV d dng acid amin, v cc VSV c kch thch pht trin bi cc acid amin. Nhn chung cc vi khun gy bnh, cc vi khun gy thi, v cc vi khun lactic (sng trong sa),... thng i hi phi c cung cp cc acid amin c sn. Cn cc loi vi khun sng trong
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t (Azotobacter, Clostridium pasterianum,...), cc vi khun t dng ho nng c kh nng t tng hp c tt c cc acid amin cn thit. Nm mc, nm men v x khun thng khng i hi cc acid amin c sn, nhng s c mt cc acid amin trong mi trng dinh dng s tng cng tc pht trin ca chng ln rt mnh m. Tuy nhin, nu mc ch nui cy l sn phm th cp, th hm lng cc cht dinh dng nit hu c phi c ti u ho i vi mc tiu sinh tng hp hot cht; v ti u hm lng sinh khi VSV trong nhiu trng hp khng song trng cng vi ti u hm lng hot cht.
1.3. Ngun dinh dng khong ca VSV

Trong cc mi trng c ngun gc t nhin (khoai ty, nc tht, sa, huyt thanh, pepton, dch chit malt,...), k c trong cng nghip, nui cy VSV, thng c vi t l thch hp cc nguyn t khong cn thit cho VSV, do vy khng cn thit phi b sung cc nguyn t khong. Nhng khi s dng cc ho cht tinh khit ch to mi trng tng hp, cn thit, phi b sung cc nguyn liu khong cn thit. Nhng nguyn t khong thit yu m VSV i hi phi c cung cp vi khi lng ln, c gi l cc nguyn t i lng. Cn nhng nguyn t khong m VSV ch i hi vi khi lng nh, c gi cc nguyn t vi lng. Thnh phn mi trng c th tnh ton sao cho nng cc cation v cc anion l hp l cho pht trin ca VSV. P l nguyn t quan trng nht trong s cc nguyn t khong cn thit cho t bo VSV (nhiu khi P chim n 50% so vi tng s cht khong). P c mt trong cu to ca nhiu thnh phn quan trng ca t bo (acid nucleic, phosphoprotein, phospholipid, nhiu coenzym quan trng nh AMP, ADP, ATP, UDP, UTP, XTP, NAD, NADP, FAD,...; trong mt s vitamin nh vitamin B12, vitamin B6, v.v,...). m bo ngun dinh dng phospho, ngi ta s dng cc loi phosphat v c nh K2HPO4, KH2PO4,... Tuy nhin, hm lng P qu cao s lm gim st nghim trng, thm ch nh ch sinh tng hp hot cht mc tiu, mc d VSV pht trin rt mnh. V d, i vi cc VSV c kh nng sinh tng hp khng sinh, cc khong nng phosphat di y cho s liu ng tham kho: + 1mmol Pi/dm3 cho php sinh tng hp ti a sn phm th cp. + 10mmol Pi/dm3 c ch ti a sinh tng hp sn phm th cp. + Cn vi nng phosphat v c l 2,5mmol/dm3 thng cho t sn phm. S l nguyn t khong quan trng trong t bo VSV. S c mt trong mt s acid amin (cystein, cystin, methionin,...), mt s vitamin (biothin, thiamin,...). Tripeptid glutation c vai tr quan trng trong cc qu trnh oxy ho kh. Phn ng chuyn sulfidrin thnh disulfid trong qu tnh chuyn in t t nguyn liu h hp n oxy phn t. a s VSV s dng S di dng mui sulfat, mt s t s 2 dng di dng thiosulfat (S 2 O3 ,...) hoc dng kh (H2S,...). Mg l nguyn t c VSV i hi vi lng khng cao (104 103M), nhng vai tr ca magie li khng nh. Magie ng vai tr mt ng tc nhn tham gia vo nhiu phn ng enzym c lin quan n cc qu trnh phosphoryl ho. Mg2+ c th lm hot ho cc enzym hexokinase, ATPase, pirophosphatase, phosphoglucomutase, carboxylase,..., cc enzym trao i protein, cc enzym oxy ho kh ca chu trnh Krebs (khong hn 80 enzym khc nhau). Mt khc magie cng ng vai tr quan trng trong lin kt cc tiu phn ribosom vi nhau. Ca l nguyn t t tham gia vo vic xy dng nn cc hp cht hu c m ng vai tr cu ni trung gian gia cc thnh phn quan trng ca t bo (gia ADN v protein trong nhn, gia cc nucleotid, gia ARN v protein trong ribosom,...). Calci cn thit cho vic hnh thnh cu trc khng gian t bo. Zn cng l ng tc nhn trong cc phn ng enzym xc tc, c tc dng ng k trong hot ho mt s enzym nh carboanhydrase, phosphatase kim, pirophosphatase, leucitinase,... Mn c cha trong mt s enzym h hp. Mn ng vai tr quan trng trong hot ho mt s enzym nh phosphomonoesterase, carboxylase, ATPase, hydroxylamin reductase, arginase, aminopeptidase, enolase, phosphoglucomutase,... Vic hot ho enzym i khi khng mang tnh c hiu, v d, isocitratase t P. aeruginosa c th
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c hot ho t cc ion khc nhau (Mg2+, Mn2+, Fe2+ hoc Co2+). C khi ion ny tc dng ngc vi ion kia: Na+ c ch pht trin ca Lactobacillus casei, nhng nu b sung thm K+ th tc dng c ch y s mt. C mt s nguyn t cho n nay vai tr sinh l ca chng vn cha c hiu r lm, trong c kali, natri, clo. K chim t l kh cao trong thnh phn ca t bo VSV nhng cho n nay vn cha thy kali tham gia vo bt k thnh phn no ca nguyn sinh cht, cng nh cha tm thy bt k enzym no c cha kali, m thy kali thng tn ti trong dng ion K+ mt ngoi ca cu trc ca t bo. Nhiu nghin cu s dng K40 cho bit, mt phn ng k kali tn ti trng thi lin kt ho l khng bn vng vi protein v cc thnh phn khc ca nguyn sinh cht. Kali c th tc dng nh cc ion kim loi khc thng qua vic nh hng n tnh cht ho keo v hot ng xc tc ca enzym. Nhiu th nghim tin hnh thay th kali trong cc mi trng dinh dng bng cc ion kim loi kim ho tr mt khc nh Na, Li, Rb, Cs,... u khng c kt qu. Tuy nhin kali cng tham gia vo vic hot ho mt s enzym (amylase, acetyl CoAsynthetase, ATPase,...). Kali lm tng ngm nc ca cc h thng keo do nh hng n cc qu trnh trao i cht, nht l cc qu trnh tng hp nh tng hp thiamin,... v c nh hng nht nh n h hp ca t bo VSV. Na v Cl cng l cc nguyn t m nhiu VSV cng cn vi s lng khng nh, nhng cho n nay ngi ta cng cn bit t v vai tr sinh l ca chng. Hm lng hai nguyn t trn c bit cao cc VSV a mn (Halophyl) sng trong nc bin, t vng ven bin, hoc pht trin c trn cc loi thc phm p mn. Bnh thng khi nui cy VSV, ngi ta khng cn b sung cc nguyn t vi lng v nhng nguyn t y thng c sn trong nc my. Tuy nhin, trong mt s trng hp ring, ngi ta vn phi b sung mt s nguyn t vi lng nht nh vo mi trng: V d: B sung Co vo mi trng nui cy VSV sinh tng hp vitamin B12; b sung Mo vo mi trng nui cy VSV c nh nit,...
1.4. Cc cht sinh trng

Cc cht sinh trng l nhng cht hu c cn thit cho hot ng sng, sinh trng pht trin i vi mt s loi VSV m khng t tng hp c chng t cc cht khc. Nhng cht c coi l cht sinh trng ca loi ny hon ton khng phi l cht sinh trng i vi loi khc, v hu nh khng c cht no l cht sinh trng chung cho tt c cc loi VSV. Ngay cng mt loi VSV, nu nui cy trong cc iu kin khc nhau cng c th c cc nhu cu khc nhau v cc cht sinh trng. V d, nm mc Mucor rouxii cn biotin v thiamin cho pht trin k kh, nhng nu nui cy trong iu kin i kh th li khng cn hai cht . iu kin pH, nhit ca mi trng, s c mt ca mt s cht dinh dng cng c nh hng r rt n nhu cu cht sinh trng ca VSV. Chng hn, nhu cu acid pantotenic ca mt s VSV (vi khun bch hu C. diphteriae) c th tho mn c khi ch cn cung cp alanin cho chng. Cc cht sinh trng cho cc VSV c th l cc nhm hp cht di y: Cc acid bo mch carbon di bo ho hay cha bo ho, chng hn C14 hoc C16,... Cc base purin v pyrimidin cng nh cc dn cht ca chng, v d cc nucleozid hay cc nucleotid,... Cc vitamin, v mt s acid amin: vitamin B12, alanin,.... y l cc hp cht m vi mt lng tuy nh nhng c tc dng quyt nh ln pht trin ca loi VSV no .
2. THNH PHN T BO V DINH DNG CA VI SINH VT

Nhu cu dinh dng ca VSV nhm p ng cc i hi ca t bo do ph thuc vo thnh phn ho hc cc t bo ca chng. Trong t bo VSV vt cht c phn thnh 2 nhm ln: Nhm th nht l nc v cc mui khong, v nhm th hai l nhm cc cht hu c (bng 2.1) bao gm acid nucleic, protein, hydratcarbon, lipid v cc cht hu c phn t nh.
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V tng th, nc chim t l cao nht trong s tt c cc thnh phn, cn trong s cc cht hu c th protein chim t l cao nht. Xt chi tit v cc nguyn t, t bo VSV bao gm cc nguyn t sinh hc C, O, N, H v cc nguyn t khong a lng v vi lng. Ch ring cc nguyn t C, O, N, H, P, S, K v Na chim n 98% khi lng sinh khi kh ca t bo E. coli (bng 2.2).
Cc s liu trn l nhng gi tr trung bnh, v trong cc iu kin sinh trng khc nhau, cng nh trong cc giai on sinh trng, pht trin khc nhau thnh phn cc t bo VSV khng ging nhau.
2.1. Thnh phn nc v mui khong

Nc chim n 70 90% khi lng t bo VSV. Cc phn ng sinh ho xy ra trong t bo VSV u din ra trong dung dch nc. vi khun, hm lng nc thng l 70 80%, cn nm si thng l 85 90%. Phn nc c th tham gia trong cc qu trnh trao i cht ca VSV c gi l nc t do. a phn nc trong t bo VSV tn ti di dng nc t do. Nc kt hp l nc lin kt vi cc hp cht hu c cao phn t trong t bo (protein, lipid, hydratcarbon). Nc lin kt mt kh nng ho tan v lu ng. Ly mt nc hay sy kh thc phm l phng php ngn cn s pht trin ca VSV, bo qun thc phm, thuc nam,... c lu hn. Mt khc p mui hoc tm ng nhm to ra s kh cn sinh l khng thch hp cho s pht trin ca VSV bo qun thc phm hay c p dng. Mui khong chim khong 2 5% khi lng kh ca t bo. Trong t bo chng thng 2 2 dng ion: dng cation nh Mg2+, Ca2+, K+, Na+,... hay dng anion nh HPO 4 , SO 4 , HCO3 , Cl ,... Cc ion trong t bo VSV lun tn ti nhng t l nht nh nhm duy tr pH thch hp cho tng loi VSV.
2.2. Cc cht hu c
Trong t bo vi khun, cc polyme cc cht i phn t chim ti 95,5 96% cht kh, 3,5% l cc cht n phn t v ch cn 0,5 1,0% l cc ion v c. Cc polyme ch yu trong t bo l cc protein, acid nucleic, lipoid, hydratcarbon. Cc cht hu c trong t bo VSV c to nn ch yu
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bi cc nguyn t C, H, O, N, P, S,... Ring 4 nguyn t C, H, O, N chim ti 9293% cht kh ca t bo. Protein sinh cht c hm lng cao nht trong t bo c to thnh ch yu bi cc nguyn t: C (50 55%), O (21 24%), N (15 18%), H (6,5 7,3%), S (0 0,24%), v lng nh mt s nguyn t khc nh P, Fe, Mg, Ca, Cu, Mn,... Cc acid amin kt hp vi nhau trong lin kt peptid to thnh cc protein,... Bng 2.3 gii thiu thnh phn ho hc ca t bo vi khun.
Vi:
(1). Khi lng kh ca 1 t bo E. coli sinh dng l 2,8 1013g, (2). Peptidoglycan v protein, (3). l 4 loi phospholipid, mi loi c nhiu nhm ph khc nhau.
3. C CH VN CHUYN CC CHT DINH DNG VO T BO VI SINH VT

tn ti, sinh trng v pht trin, t bo VSV phi thng xuyn trao i cht v nng lng vi mi trng xung quanh. Mt mt chng nhn cc cht dinh dng cn thit t mi trng, mt khc th thi ra mi trng bn ngoi cc sn phm trao i cht. Gia mi trng xung quanh v t bo cht trong t bo tn ti hng ro thm thu, hng ro ny chnh l mng t bo cht lipoprotein: cc hp cht a lipid (lipofil) xm nhp t bo nhanh hn cc hp cht k lipid (lipofob). Chng minh kt lun trn, cc nh nghin cu x l VSV vi cc dung mi lipid nh nbutanol lm hng ro thm thu ca VSV b hu hoi gy ra vic tch thot cc cht phn t thp ra khi t bo. Nh vy mng t bo cht l c quan m bo khng gian v iu chnh c chn lc s chuyn vn vo, ra ca cc cht khc nhau. S xm nhp ca nc v cc cht ho tan qua mng t bo cht l mt qu trnh ng hc theo mt trong hai c ch: khuch tn n gin hay khuch tn th ng v vn chuyn ho lp th c hiu. Trong khuch tn th ng, cc phn t i qua mng t bo cht mt cch n gin nh vo mc chnh lch nng (trong trng hp cc cht khng phn ly), hay chnh lch in th (trong trng hp cc ion). Cc nghin cu cho thy, ngoi nc, ch mt s t hp cht c chuyn vn theo c ch ny. Trong vn chuyn c hiu nh protein thm (a s cc cht c vn chuyn qua mng t bo theo c ch ny), cc phn t vn chuyn (cc protein vn chuyn hay cc protein thm cc permease) sp xp trn mng lin kt vi cc cht dinh dng ri chuyn chng vo b mt bn trong ca mng, m t y chng c chuyn vo t bo cht. Kiu vn chuyn cc cht dinh dng nh vy c th l th ng (khng cn nng lng ca t bo vn chuyn xui dng). Phc cht cht ho tan permease c vn chuyn theo c hai pha nh chnh lch nng ca cht no . C ch c bn y c th lin quan n i lc gn kt ca protein vn chuyn vi cht tan cn vn chuyn hai pha trong v ngoi mng t bo cht l khc nhau: nu i lc ca cht mang vi cht tan bn ngoi mng cao hn bn trong,
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cht dinh dng c vn chuyn t mi trng ngoi vo trong t bo, cn i vi cht thi th ngc li. Nhng vn chuyn cc cht m nng trong t bo cao hn ngoi mi trng, tc vn chuyn cht phi thc hin ngc vi gradient nng , theo kiu ngc dng, cn i hi tiu th nng lng, v nng lng tiu hao c cung cp t ATP (c hnh thnh trong mesosom hoc mng t bo cht). Cc nghin cu cho thy c ATPase (adenosin triphosphatase) trong mng t bo cht ca cc vi khun, l enzym c lin quan n vn chuyn cc cht. Trong vn chuyn ch ng, enzym vn chuyn (Pa) dng hot ng gn vi cht tan (S) cn vn chuyn pha ngoi mng to thnh phc hp cht tan permease (S P). Phc hp ny c di chuyn vo pha trong mng v c ATP cung cp nng lng, bin thnh dng permease (Pi) t do bt hot pha trong mng t bo cht, ci bin cu trc, gii phng cht tan vo t bo cht. Lp tc Pi di chuyn ngc chiu ra pha ngoi mng v li c bin dng thnh Pa hot ng, chu k vn chuyn c lp li. Chng hn K+ c th c vn chuyn theo c ch ny. Vic thay i cu trc Pi Pa c lin quan n s thay i cu trc bc ba ca protein vn chuyn (hnh 2.1).
1) Khuch tn n gin
Ngoi mng Trn mng Trong mng
2) Chuyn vn th ng
Hi phc Kt hp Ci bin
3) Chuyn vn ch ng
Hi phc Kt hp Ci bin + E
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Hnh 2.1. Vn chuyn ch ng cht dinh dng vo t bo
Kiu vn chuyn ch ng khc l kiu vn chuyn kt hp phosphoryl ho, c lin quan n vn chuyn cc ng (v d: glucose, mannose,...): bc th nht xy ra trong pha t bo cht, protein vn chuyn c phosphoryl ho kt hp vi bin i phosphoenolpyruvat. Phc hp cht mangphosphat (PMPr) di chuyn ra pha ngoi mng, v lin kt vi glucose cn vn chuyn thnh phc hp GluMPr, vn chuyn ng vo t bo cht v gii phng ng phosphoryl ho vo t bo cht (hnh 2.2). y phn ng phosphoryl ho c thc hin nh enzym phosphattransferase EI v EII. EI hot ng pha bn trong v EII hot ng pha bn ngoi mng.
Trong Mng CM Ngoi
ngP
P enolpyruvat
MPr
Pyruvat
PMPr
ng
Hnh 2.2. Vn chuyn kt hp phosphoryl ho
Tnh c hiu c cht ca cc protein (enzym) vn chuyn rt khc nhau: mt s c tnh c hiu cao, v d: permease ca galactose ca E. coli ch cho php vn chuyn galactose. Tnh c hiu ca cc permease ca cc ng khc v cc acid amin c tnh c hiu thp hn. Cn nhn mnh rng, theo m hnh hnh 2.1 th cng mt permease c th m nhn c chc nng vn chuyn th ng ln ch ng (tu theo s vng mt hay c mt ca ATP) v phn ng hot ho c cung cp nng lng nh ATP din ra pha trong ca mng, tuy nhin y cng ch l gi thit.
CC CU HI LNG GI
1. Trnh by cc nhm dinh dng carbon VSV. 2. Trnh by cc cht dinh dng chnh ca VSV. 3. Trnh by cc kiu vn chuyn cht dinh dng t bo VSV.
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Chng 3
TRAO I CHT V TRAO I NNG LNG
1. CC KHI NIM CHUNG

Trao i cht l qu trnh hp th cht dinh dng t mi trng qua qu trnh d ho phn gii chng, gii phng v tch lu nng lng v to ra cc cht trung gian quan trng, v t y qua ng ho tng hp ra cc cht cn thit cho sinh trng, pht trin ca t bo. Nh vy, trao i cht bao gm hai mt d ho v ng ho b sung cho nhau. VSV hp th cht dinh dng v qua qu trnh chuyn ho phn gii cc cht dinh dng thnh cc cht trung gian, ng thi tch lu nng lng c gii phng trong cc phn t cha nng lng cao y l qu trnh d ho. Mt khc, qu trnh ng ho l qu trnh tng hp cc thnh phn c th v gii phng cc sn phm trao i cht (cc cht chuyn ho). D ho v ng ho l hai mt b sung cho nhau gip cho VSV thc hin c chc nng sng ca mnh trong quan h trao i cht v nng lng vi mi trng xung quanh. Xt t gc nng lng, cc qu trnh oxy ho phn gii c cht km theo gii phng nng lng c gi l qu trnh trao i nng lng. ng vt v thc vt bc cao cc qu trnh ny c th xy ra vi cc c cht dinh dng thng nht (cc cht hp th t mi trng) v cc cht d tr ca c th (protein, polysaccharid, lipid). Nhng t bo VSV, s lng cc cht d tr thng rt nh v th chng phi s dng ch yu l cc cht hp th c t mi trng xung quanh l chnh. Phn ln cc loi VSV thuc loi dinh dng ho nng, chng chuyn ho cc hp cht ho hc lm ngun to ra nng lng. Nng lng mi gii phng c ny ch mt phn nh bin thnh nhit nng, cn phn ln truyn t hp cht ny sang hp cht khc cng vi cc in t hoc cc nguyn t. Cc VSV d dng ho nng hay dinh dng ho nng hu c s dng cc hp cht dinh dng hu c, cn cc VSV t dng ho nng hay dinh dng ho nng v c th li s dng cc hp cht dinh dng v c. Cc VSV d dng quang nng bao gm nhm dinh dng quang nng v c v nhm dinh dng quang nng hu c s dng trc tip nng lng ca nh sng vi cc h thng bin i nng lng thch hp. Cc VSV i kh s dng trc tip oxy khng kh kt thc qu trnh oxy ho nu. Cn vi cc VSV k kh, qu trnh oxy ho sinh nng lng khng kt thc bng vic kt hp vi oxy khng kh m vi cc dng oxy hp cht theo cc hnh thi oxy ho khc: bn cnh vic lin kt vi oxy qu trnh oxy ho cn l s mt bt hydro (RH2 + A R + AH2), qu trnh tch hydro sau khi kt hp vi nc (A + H2O B C), v qu trnh mt bt in t (e) lm gia tng s ho tr dng (Cu+ Cu2+ + e, Fe2+ Fe3+ + e). Nng lng gii phng ra t cc phn ng oxy ho c tch lu trong cc cht giu nng lng, cc nucleozid triphosphat: GTP, ATP, UTP; cc acyl
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phosphat; cc dn xut acid carbonic (AcetylCoA,...). Hp cht giu nng lng bn vng nht, quan trng nht l ATP (adenosin triphosphat xem cng thc di). ATP cha 2 lin kt nng lng cao (~). Trong 1 mol ATP, mi lin kt cao nng cha khong 11.500cal, trong khi lin kt thng ch cha 300cal. ATP c coi l kho cha ca nng lng trong t bo, c s dng trong tt c cc phn ng cn nng lng. Cc phn t ATP giu nng lng ny c to ra trong t bo VSV cng nh trong t bo ca mi sinh vt khc. Qu trnh to ra ATP trong cc c th Procaryota khng quang hp c thc hin theo hai c ch chung bao gm: qu trnh phosphoryl ho c cht v qu trnh phosphoryl oxy ho, l phng thc ch yu tch ly nng lng cho t bo. Trong c hai trng hp, t ADP v phosphat v c s dng nng lng lin kt ho hc ca cc sn phm trao i cht trung gian m sn xut ATP. Trong qu trnh th nht, nng lng t lin kt cao nng ca c cht, v d: 1,3 diphosphoglycerat hay phosphoenolpyruvat chuyn sang cho ATP. Qu trnh th hai xy ra khi oxy ho NADH2 hay NADPH2,...
NH2 N N N N
H CH2 O
O P OH
O OH
O OH
O ~ P O ~ P OH
OH H
O H
adenin
H O ribose adenosin AMP ADP AT P
phosphat phosphat phosphat
2. CC CON NG OXY HO SINH HC PHN GII HYDRATCARBON CC VI SINH VT D DNG

Oxy ho sinh hc l khi nim chung cp n hng lot cc phn ng oxy ho hydratcarbon sinh nng lng xy ra trong cc t bo sng. Oxy ho sinh hc khc vi s chy ch: phn ng xy ra nhiu bc, trong iu kin n ha, c xc tc l cc loi enzym, nng lng phn ln c tch lu trong ATP v hiu sut s dng nng lng cao.
2.1. Phn gii glucose

Glucose l ngun vt cht v nng lng cho cc t bo VSV. Sau khi c hp th vo trong t bo, glucose c chuyn ho qua cc hp cht trung gian cha mch carbon khc nhau theo 3 con ng ring bit n acid pyruvic (pyruvat) sn phm cui chung ca cc con ng ng phn. Phn t glucose kh tr v mt ho sinh, do trc tin glucose c hot ho nh phosphoryl ho v tr C6 chuyn thnh glucose6phosphat l dng hot ng chuyn ho
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trong t bo v l im chung ca c 3 con ng ng phn. 2.1.1. Con ng ng phn (glycolysis) Con ng phn gii ng ny cn c gi l con ng EMP (EmbdenMayerhofParnas pathway) (hnh 3.1).
Hnh 3.1. Con ng ng phn Embden-Mayerhof-Parnas
Trong iu kin c hay thiu vng oxy, glucose u c chuyn ho thnh pyruvat qua 10 phn ng, trong ch c 3 phn ng (1, 3 v 10) l mt chiu, cn li l cc phn ng thun nghch. Tr nguyn liu xut pht (glucose) v hp cht cui (pyruvat), cc hp cht trung gian u dng phosphoryl ho. Gc phosphoryl y c 3 chc nng: Gip cho hp cht trung gian mang in tch m cao khng th thot ra ngoi. L v tr gn enzym.
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L v tr cung cp lin kt phosphat cao nng (~P). Phn ng tng qut:
y, ATP c to thnh trong 2 bc (trong tng s 3 bc to nng lng) phn ng: th 7 v th 10. c chun b t bc th 6 vi vic to thnh phn t cao nng NADH2, bc phn ng th 7 l bc kt thc ca phn ng oxy ho:
Enzym xc tc phn ng 6 (3Pglyceraldehyd dehydrogenase) cha nhm SH, do b km hm bi cc kim loi nng. Mt khc 1,3bisPglycerat c cha mt ~P C1. Vi s hin din ca 3 3 gc arsenat AsO 4 do cu trc tng t vi gc PO 4 , xy ra s cnh tranh to thnh lin kt ~P C1, dn n vic km hm s to thnh ATP. Cn bc phn ng 10 l bc kt thc ca bc phn ng loi nc th 9:
S to thnh ATP hai phn ng trn l phosphoryl ho c cht v phosphat cao nng (~P) trc l mt phn ca hp cht trung gian (1,3bisPglycerat v PEP). Khi c mt ADP, cht chuyn ho s chuyn ~P cho ADP:
Con ng ng phn EMP cung cp cho t bo 6 trong s 12 tin cht tng hp cc n v kin trc: Glucose6P, Fructose6P, 3Pglyceraldehyd, 3Pglycerat, PEP, v pyruvat. V nng lng, con ng EMP sinh li 2 ATP v 2NADH2. NADH2 l phn t cao nng, cha 52.000cal/mol. Tuy nhin v khng bn vng bng ATP, nn NADH2 s truyn nng lng cho ATP trong chui h hp sau ny. T phn ng s 6 n phn ng s 10 ca con ng ng phn ny c gi l giai on phn gii cui (GPGCterminal glycolytic pathwayTGP). 2.1.2. Con ng pentosephosphat (PP hay con ng hexosemonophosphat HMP) Cng nh EMP, trc tin glucose c phosphoryl ho thnh glucose6P v qua hai phn ng c xc tc bi glucose6P dehydrogenase v 6Pgluconatdehydrogenase, glucose6P c bin i thnh ribulose5P v gii phng CO2. Qu trnh oxy ho c tip tc thng qua cc phn ng tip theo: mt mt l cc phn ng chuyn ho qua li gia pentoseP v hexoseP, t con ng ng kn thnh chu trnh sau mi mt chu trnh th mt CO2 c ct ra kt hp vi gii phng tch lu nng lng, mt khc l glyceraldehyd3P i vo giai on phn gii cui GPGC ca con ng ng phn v c bin i thnh pyruvat. Con ng phn gii ny gip cho vi khun chuyn ho glucose thnh pyruvat khng qua EMP, ng thi cung cp cho t bo cc hp cht trung gian khc C3 v C6 nh l ribose5P ( tng hp acid nucleic) v erythrose4P (cng vi PEP), cng nh sedoheptulose7phosphat tng hp cc acid amin thm, cc nhn t cu trc cng nh chc nng khc ca t bo,...
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ng thi HMP cn cung cp NADPH2 v NADH2 cn cho cc phn ng tng hp kh ho (hnh 3.2). Phn ng tng qut ca con ng HMP ny l:
Qua con ng trao i cht ny, phn t glucose c mc oxy ho cao nht: 1/2 phn t b t chy thnh CO2, kt hp gii phng nng lng tch lu trong cc phn t cao nng NADPH2 v NADH2 khng bn vng (ch 1ATP c trc tip to ra) v s c bin i thnh ATP trong giai on chuyn ho tip theo. (Ch rng NADPH2 hoc NADH2 cha 55kcalo/mol trong khi 1mol ATP cha 22kcalo). 2.1.3. Con ng 2keto3deoxy6Pgluconat (KDPG, 2keto3deoxy6phosphatgluconat pathway) con ng ny cn c gi l con ng EntnerDoudoroff (EDEntnerDoudoroff pathway) (hnh 3.3).
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Hnh 3.2. Con ng hexosemonophosphat (HMP), vi: G6PDH Glucose6phosphatdehydrogenase v 6PGDH 6phosphogluconatdehydrogenase.
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CH2OH H H O O H H OH H H OH OH
ATP
2+
ADP H
CH2O P H OH
NADPH2 O H NADP H
CH2O P O H OH
Mg
Glucose
O H G-6-PDH H O OH OH OH H H 6-Phosphoglucono-1,5-lacton Glucose-6-phosphat H O H H2O, Mg

2+
Lactonase COOH C O H C H hoc H H O
CH2 O OH H H H HO
P O
H C OH H C OH
CH2OPO3
CH2O P OH Dehydratase H HO H O 2+ Fe , GSH H OH H O H2O OH H 6-Phosphogluconat
2-Keto-3-deoxy-6-phosphogluconat Aldolase
O COOH C O CH3 Pyruvat CH C O CH2 O P Pyruvat
Glyceraldehyd-3-phosphat
Hnh 3.3. Con ng EntnerDoudoroff (ED)
Trc tin glucose c phosphoryl ho thnh glucose6phosphat, v glu6P c chuyn thnh 6Pgluconat nh trong con ng PP. Nh tc dng ca phosphogluconatdehydratase, 6P gluconat b mt nc to thnh 2keto3deoxy6phosphatgluconat (KDPG). KDPG ny b phn gii thnh pyruvat v 3Pglyceraldehyd nh aldolase c hiu. Cui cng 3Pglyceraldehyd li i vo giai on phn gii cui ca glucoseGPGC v c bin i thnh pyruvat. Phn ng tng qut ca con ng chuyn ho ny l:
Con ng phn gii glucose ny cng c pht hin trong mt s vi khun nh l con ng chuyn ho chnh. Cc VSV khc nhau s dng 3 con ng phn gii glucose vi mc khc nhau (bng 3.1). Con ng EMP v PP ph bin nhiu VSV, cn con ng KDPG gip cho nhiu vi khun s dng c gluconat.
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E. coli, S. cerevisiae, P. chrysogenum,... v nhiu loi Clostridium ng phn glucose qua EMP, nhng phn gii gluconat qua con ng KDPG. Pseudomonas aeruginosa phn gii glucose qua HMP v trao i gluconat qua KDPG. Mt s Clostridium v vi khun hiu kh chuyn ho gluconat qua con ng KDPG ci bin: gluconat c chuyn thnh 2keto3deoxygluconat nh gluconatdehydratase v ch bc ny mi c phosphoryl ho s dng ATP nh ketodeoxygluconokinase; KDPG b phn gii nh 2keto3deoxyPgluconat aldolase,.... Tng qut li sn phm cui cng ca cc con ng ng phn l: EMP: Glucose 2 pyruvat. HMP: Glucose 1 pyruvat + 3CO2. ED: Glucose 2 pyruvat.
2.2. Phn gii cc loi ng khc

Ngoi glucose, VSV cn c th s dng c cc loi ng khc nhau. Cc loi ng n trc tin u phi c phosphoryl ho, ri bin i thnh cc dn cht tng ng m i vo con ng ng phn thch hp. Fructose sau khi c phosphoryl ho s i vo EMP:
Fructokinase
Fructose
Fructose-6-phosphat
Mannose sau khi c phosphoryl ho nh mannokinase thnh mannose6P, sau c phosphomannoseisomerase bin i thnh fructose6P v i vo EMP:
Mannokinase Phosphomannoseisomerase
Mannose
Mannose-6- P
Fructose-6-P
Galactose sau khi bin thnh Glucose6P cng i vo EMP:
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Galactokinase
Galactose
Galactose-1- P
Phosphoglucose uridin transferase
Glucose-1- P
Phosphoglucose mutase
Glucose-6- P
Pentose sau khi c phosphryl ho s i vo HMP. Cc trisaccharid v olygosaccharid s c VSV thu phn thnh cc monosaccharid trc khi s dng. Mt s VSV c kh nng phosphoryl ho trc tip disaccharid. V d Lactobacillus v Neisseria meningitidis c enzym maltophosphorylase c th xc tc phn ng:
Nh vy, trong trng hp ny khng cn s dng nng lng t ATP m vn c th to ra c glucose1phosphat.
3. QU TRNH OXY HO PYRUVAT V CHU TRNH KREBS (CHU TRNH TRICARBOXYLIC ACIDTCA)
3.1. Qu trnh oxy ho pyruvat

Pyruvat ng vai tr trung tm trong trao i cht trung gian, l tin cht ca nhiu sn phm chuyn ho. Nhiu VSV oxy ho pyruvat thnh acetylCoA. C 3 phn ng quan trng i vi cc VSV:
PDH
+ 1. Pyruvat + CoA + NAD
AcetylCoA + NADH2 + CO2
2. Pyruvat +
CoA +
2Fd
PFdOxr.
Acetyl-CoA + 2FdH
CO2
3. Pyruvat +
CoA
PFL
Acetyl-CoA
Format
Vi: Fd = ferredoxin. Phn ng 1 do phc h pyruvatdehydrogenase (PDH) xc tc. PDH c VSV hiu kh (khng c VSV k kh). AcetylCoA mi to ra i vo chu k TCA. PDH gm c 3 phc h enzym. iu kin cho phn ng ngoi CoA v NAD+ cn cn TPP (thiaminpirophosphat) v acid lipoic (hnh 3.4). Phn ng 2 c xc tc bi pyruvatferredoxinoxydoreductase (PFdOxr), gp nhiu vi khun k kh nh Clostridium. Phn ng 3 do pyruvatformatliase (PFL) xc tc, enzym ny c trong cc vi khun k kh sinh acid formic (h Enterobacteriaceae) cng nh vi khun quang dng.
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CH3
COO + + (+) + H + H + () C C O () COO CH3 C OH CO2

- +
carboxylase
carboxylase
H CH3 C C carboxylase OH acetaldehyd hot ng
CH2 CH2 S SH CH2 CH2 HOOC O (CH2)4 CH S C CH3
CH S acid lipoic O C (CH2)4 Enzym
S-acetyldihydrolipoic acid CoA-SH CoA S C CH3 O CH2 CH2 SH HOOC (CH2)4 CH SH dihydrolipoic acid NAD
+
NADH2
Hnh 3.4. Oxy ho pyruvat nh pyruvat dehydrogenase
3.2. Chu trnh Krebs

AcetylCoA to ra nh trn c oxy ho trit qua chu trnh Krebs gii phng CO2, nc v tch lu nng lng (hnh 3.5).
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CH3 C O COOH
CoASH NAD
NADH 2 O CH C 3 CO2 Acetyl-CoA S CoA
Pyruvat
H C COOH 2 Citratsynthase H C COOH Citrat O H C COOH 2 Aconitase HO 2 H C COOH 2 C COOH H C COOH HO 2
O Oxaloacetat NADH 2 NAD

+
C COOH H C COOH 2 Cis-aconitat Malat-DH
Aconitase
H Malat H C COOH O H C COOH 2 Fumarase
H C COOH 2 H C COOH Isocitrat H C COOH O H i-Citrat-DH Oxalosuccinat NADP

+
HO 2
NADPH 2
H OC C H O FumaratH C COOH FADH 2 FAD

+
H C COOH 2 H C COOH O C COOH
i-Citrat-DH Succinat-DH CO2
Alfa-ketoglutarat H C COOH 2 C COOH H 2 Succinat CH 2 H C COOH 2 H C COOH CO2 2 AK-DHO C COOH SCoA-TK CH 2 CoASH O C SCoA + CoASH ADP NAD Succinyl-CoA ATP + Pi NADH 2
Hnh 3.5. Chu trnh Krebs
Chu trnh Krebs oxy ho acetat gii phng CO2 v tch chuyn H2. CO2 c tch ra trong 2 bc nh isocitratdehydrogenase v ketoglutaratdehydrogenase. Trong s 8[H] c tch khi acetat th 6[H] c chuyn n cho NAD+ v NADP+ nh 3 dehydrogenase (DH) l i citratDH, ketoglutaratDH v malatDH, cn 2[H] c chuyn n cho FAD+ nh succinatDH. Chu trnh TCA cung cp cho t bo nh oxaloacetat, ketoglutarat tng hp acid amin (aspartat v glutamat) v succinylCoA s dng trong tng hp Hem. Nng lng sinh ra ch yu c gi trong cc phn t cao nng l cc cht kh 3NADH2, 1NADPH2, 1FADH2, cng nh 1ATP. Kh nng gi nng lng trong cc hp cht ny khng bn nh trong ATP v nu khng c s dng gia chng th n giai on cui cng ca chui h hp s chuyn sang cho ATP. Phng trnh tng qut ca chu trnh Krebs t pyruvat l:
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Nng lng tch lu trong cc sn phm phn ng l:
Phn ng 2 pyruvat + 2H+ + 5O2 6CO2 + 4H2O c bin i entalpi t do l 273 kcalo/mol. Nh vy 90% nng lng gii phng trong t chy acid pyruvic c mt trong cc sn phm phn ng, v 75% nng lng ca glucose c chuyn sang cho cc sn phm hu ch.
4. CHU TRNH GLYOXYLAT

Khi VSV sinh trng trn acetat hoc cc hp cht m phn gii dn n acetat (cht bo, hydrocarbon,...) din ra chu trnh glyoxylat (chu trnh KrebsKornberg). Chu trnh ny xy ra tng t nh chu trnh Krebs, nhng do khng c enzym ketoglutarat dehydrogenase m thiu mt mt s bc phn ng t chu trnh Krebs v i qua hai phn ng chnh:
ln lt do isocitratliase v malatsynthase xc tc. Nng lng t c trong chu k ny khng cao: 1NADH2, nu tnh c chu k b tr mi thm 1NADH2 na (hnh 3.6). Chu trnh glyoxylat cng bin i oxaloacetat thnh pyruvat v CO2 nh xc tc ca enzym pyruvat carboxylase hoc thnh phosphoenol pyruvat v CO2 (enzym ITP: Inosintriphosphatase). C hai hp cht triose ny, sau , u c th c bin i thnh glucose nh qu trnh EMP o ngc. Chu trnh glyoxylat c tm thy nhiu vi khun, v d nh Avinelanohi,...
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Acid bo
-oxy ho
O CH3 C S CoA
Acetyl-CoA
Citratsynthase H2 C COOH Oxaloacetat HO C COOH Citrat H2C COOH
O C COOH H2C COOH NADH2
Aconitase NAD
+
Malat-DH O HO C Malatsynthase H C
H Glucose HO C COOH H2C COOH Malat
I-citrat-liase
H2C COOH H C COOH
HO C COOH O Isocitrat H Glyoxylat Succinat CoA-SH Acetyl-CoA + +H CO2 -ketoglutarat
Fumarat
FADH2 Succinat Succinyl-CoA CO2
Hnh 3.6. Chu trnh Glyoxylat
5. CHUI H HP V PHOSPHORYL HO OXY HO KH (respiratory chain and oxidative phosphorylation)

Vi khun hiu kh tng hp ATP rt mnh m hn hn vi khun k kh nh c chui h hp v enzym ATPsynthase (yu t F0 F1). Chui h hp l h thng phc hp enzym nhiu bc c
nhim v gip cc VSV i kh oxy ho hydro v x l cc e tch lu li sau cc chuyn ho oxy ho v chu trnh Krebs, cn ATPsynthase hp tc vi chui h hp. Hai h thng ny nm mng: mng t bo cht hoc mng trong ty th. Ion H+ v in t e (electron) tch ra t c cht i vo chui h hp; cc e c chuyn n O2. Nng lng tch lu c chuyn thnh ATP l chnh, phn nh c gii phng di dng nhit. Cc thnh phn ca chui nm trong lp phospholipid kp gm nhiu enzym vn chuyn e v H+, cc coenzym v cc dehydrogenase v h thng vn chuyn. Cc thnh phn quan trng nht tham gia vo oxy ho hydro l: Flavoprotein l cc enzym cha cc coenzym FMN hoc FAD vn chuyn hydro. Phc protein Fe S vn chuyn e cha Fe: Phc protein Fe S va lin kt vi S ca cystein, va lin kt vi S ca H2S. Cystein l mt phn ca chui polypeptid, v trung tm Fe S c th c coi l nhm b sung ca chui polypeptid. Trung tm Fe S thuc typ [2Fe 2S], ch vn chuyn 1e, c cha 2Fe v S. Trong lng Fe bt gp mng t bo th n 80% nm trong
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phc protein Fe S, cn 20% nm trong cytocrom. Cc protein Fe S cng tham gia vo c nh nit, kh sulfit, kh nitrit, quang hp v gii phng, hot ho H2. Phn t lng M ca protein Fe rt thp v th oxy ho kh rt m. EO nm trong khong 0,2 0,6 V. Ngoi protein [2Fe 2S], lc lp vi khun hiu kh cn c trung tm [4Fe 4S] (Chromatium, Azotobacter,...). Mt s protein [2Fe 2S] quen thuc: Ferredoxin, putidaredoxin, rubredoxin hoc adrenodoxin. Quinon: mng trong ty th v vi khun G l ubiquinon (CoQ), vi khun G+ l naphtoquinon, v lc lp l plastoquinon. Quinon v c bit ubiquinon c c tnh a lipid do gn vo lp phospholipid kp ca mng, vn chuyn hydro hoc e . So vi cc thnh phn khc ca chui th quinon c d 10 14 ln, v l kho d tr hydro chuyn vo chui h hp qua cc coenzym v nhm b sung khc nhau, ri chuyn tip hydro n cc cytocrom. Cytocrom: H thng cytocrom vn chuyn e, khng vn chuyn hydro. Cytocrom nhn e t quinon: trong qu trnh vn chuyn, mt lng H+ tng ng e b tch vo mi trng. Cytocrom cha nhm b sung l Hem. C mt s loi cytocrom: b, c1, c, a, a3, O. Cytocrom gp hu ht cc VSV c chui h hp. Cytocromoxydase (cyt. O) chuyn 4e trc tip ln oxy (hnh 3.7):
Cc thnh phn ca chui h hp c sp xp trn c s th oxy ho kh, bt u t th m nht NAD+ cho n Cyt.O. Mt s cht c tc dng ln chui h hp nh amital, rotenon,... km hm NAD dehydrogenase; antimixin A km hm gia cyt.b v c; cyanid (CN) km hm Cyt.O. Khi chuyn 2[H] qua NAD+ cho h s to ATP P/O = 3 (ATP mol/ nguyn t oxy), cn khi chuyn 2[H] qua FAD+ cho h s to ATP P/O = 2. Tuy nhin vi khun (v d: E. coli) do hn ch v tr chuyn 2[H] c h s P/O = 3, nn h hp glucose hiu kh ch cho 26 ATP thay v 38 ATP.
H2O
+
H ng phn
NAD
2-
Chu trnh Krebs
NADH2
FAD Fe3+ FADH2 Pro(2Fe2S) Fe

2+
CoQ CoQH2
Fe
3+
Cyt.b
Fe2+
Fe3+
Fe3+
Cyt.c
Fe
2+
Qu trnh oxy ho khc
Cit.a
Fe2+
Cytcrom oxydase 1 O2 2 2+
Fe
Fe
3+
ADP
ATP
Hnh 3.7. Chui h hp trong qu trnh oxy ho sinh hc
6. H HP K KH V CC QU TRNH LN MEN K KH
6.1. H hp k kh
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Trong iu kin thiu oxy, nhiu vi khun c th tin hnh h hp k kh, s dng oxy dng hp cht lm cht nhn hydro cui cng. C 4 kiu h hp k kh sau: H hp nitrat (k kh khng bt buc):
(AH2 l c cht, l cht cho in t). H hp sulfat (k kh bt buc):
H hp k kh c bit l kh CO2 vi c cht l H2:
Ln men k kh.
6.2. Ln men k kh
Ta gi qu trnh phn gii hydratcarbon trong iu kin k kh l qu trnh ln men k kh. y l qu trnh oxy ho kh c cht m kt qu l mt phn c cht b kh v phn khc b oxy ho. Oxy phn t khng tham gia vo qu trnh oxy ho ny, nn v thc cht y l qu trnh tch hydro ra khi c cht. Hydro mi tch ra c th c thi ra di dng kh, hoc li c th c lin kt ngay vi cc sn phm phn gii ca chnh cc c cht hu c . Nng lng sinh ra trong qu trnh ln men mt phn c s dng vo cc phn ng kh, mt phn tch lu trong cc lin kt cao nng. C th thy r rng, nng lng c to ra trong qu trnh ln men k kh khng th c di do nh trong h hp hiu kh. Trong qu trnh ln men ny, sn phm cao nng NADH2 c to ra khi ng phn (chu trnh EMP) s khng c chuyn n chui h hp nu m s tc dng vi acid pyruvic hoc cc hp cht mi c to ra t acid pyruvic thc hin vic ti to NAD+. Trong cc VSV k kh bt buc khng c nhiu loi enzym h hp nh: cytocrom a, b, c, cytocromoxydase, peroxydase, catalase,... Khc vi h hp hiu kh, sn phm cui cng ca cc qu trnh ln men k kh ngoi CO2 cn c cc sn phm c mch carbon cn cha b oxy ho hon ton (nh ru, mt s acid hu c, xeton, aldehyd). T acid pyruvic tr i, qu trnh chuyn ho trong cc VSV khc nhau l khng ging nhau, v tu thuc vo tng loi VSV v iu kin ln men m ta c cc qu trnh ln men c th: ln men acid lactic, ln men ru,... Bn cnh vic lm t cc mch carbon, trong mt s qu trnh ln men cn c s tng hp, tc l qu trnh gnko di thm mch carbon, v d: C. butiricum c th to ra acid butyric (4C) t cc sn phm trung gian 2C, 3C acid acetic, glycerin. Hnh 3.8 tm tt cc kiu h hp VSV, v hnh 3.9 Cc qu trnh ln men k kh chnh.
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Glucose (H) ATP Nm men Cc Lactobacillus Acid pyruvic Acetaldehyd (H) Cc Enterobacteriaceae Cc Propionibacterium CO2 Acid oxaloacetic (H) Acid succinic CO2 Acid propionic Butadiol Butanol Isopropanol CO2 Acid acetic (H) Acetyl CoA +HCOOH (H) Cc Clostridium AcetylCoA+ H2 C O2 ATP CO2 (H)
Acid lactic
Etanol
Etanol Acid acetic Etanol H2 CO2 AcetoacetylCoA Acid acetic (H) ATP Acid butyric ATP (H) CO2 Aceton (H)
Hnh 3.9. Cc qu trnh ln men k kh chnh
7. LN MEN I KH
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Ta gi cc qu trnh phn gii, chuyn ho hydratcarbon trong iu kin i kh l qu trnh ln men i kh. Nu quan nim cht ch th ch nhng qu trnh ln men sn xut cc sn phm s cp c cp nh ln men sn xut acid acetic, acid citric, acid glutamic,... Tuy nhin, nu hiu theo ngha rng, ln men i kh bao gm tt c nhng qu trnh ln men chuyn ho cc ngun carbon sinh tng hp cc sn phm s cp v th cp, trong c ln men sn xut cc khng sinh, k c ln men sn xut mt s vitamin,... l nhng qu trnh ln men m sn phm c nhng ng dng ht sc quan trng trong Y Dc hc. Tt c cc qu trnh ln men sinh tng hp khng sinh l cc qu trnh ln men i kh. Tuy nhin, so vi t khi mi a vo sn xut khng sinh (nhng nm 40 ca th k XX) th cng ngh khng sinh v vitamin c nhng thay i rt to ln v c mt s c im sau: Cng ngh khng sinh s dng cc chng vi nm, x khun, vi khun tri qua ci to ging nhiu bc c kh nng sinh tng hp hot cht rt cao so vi chng mi phn lp c ban u. Cc qu trnh ln men sn xut l cc qu trnh ln men v trng tuyt i, c th c iu khin hon ton bng my tnh in t online (computer online control hay digital controlDC).
8. S PHN GII PROTEIN (QU TRNH THI RA)

Qu trnh thi ra l qu trnh phn gii chuyn ho s dng c cht l protein (khc vi ln men s dng c cht l hydratcarbon). Protein l thnh phn quan trng ca xc ng vt, thc vt v VSV. Protein cha khong 15,0 17,6% nit tnh theo cht kh. Tng lng nit ca cc c th sng trn Tri t l khong 10 25 t tn, so vi tng lng carbon l khong 700 t tn. Trong lp t su 30cm bao quanh phn lc a ca Tri t c thng xuyn khong 3,0 7,5 t tn nit tn ti ch yu trong cc hp cht hu c cha nit. Phn gii cc hp cht hu c cha nit l mt khu quan trng ca vng tun hon vt cht trong t nhin, v c ngha rt ln i vi nng nghip. Qu trnh phn gii ny cn c gi l qu trnh amoni ho. Protein cng nh cc polyme sinh hc khc l dng khng s dng trc tip c i vi cc VSV. Do , mun phn gii protein, trc tin VSV phi tit enzym protease ngoi bo thu phn protein thnh polypeptid v olygopeptid. Cc polypeptid v olygopeptid ny; hoc tip tc c thu phn thnh cc acid amin nh cc peptidase ngoi bo ri cc acid amin y c hp th vo trong t bo; hoc cc polypeptid v olygopeptid c hp th vo trong t bo v ti y c thu phn thnh cc acid amin. Mt phn cc acid amin ny c VSV s dng lun trong qu trnh sinh tng hp cc protein ring ca chng, phn khc c phn gii theo cc con ng c th gii phng NH3, CO2 v cng nh sinh ra cc sn phm trung gian khc (hnh 3.10).
Rt nhiu loi VSV tham gia vo qu trnh amoni ho trong t nhin, trong s ng ch nht l cc loi sau: Vi khun: Cc loi Bacillus nh B. cereus, B. histoliticus, B. megaterium, B. mesentericus, B. mycoides, B. subtilis; Clostridium sporogenes, C. welchii; E. coli; Proteus vulgaris; Pseudomonas
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aeruginosa, P. fluorescens, P. putreficans,... X khun v nm: Streptomyces fradiae, S. griseus; S. rimosus; Aspergillus flavus, A. niger, A. oryzae; Mucor ssp; Picillium camemberti, Rhizopus ssp,... Trong t bo VSV, cc acid amin c chuyn ho nh loi nhm amin hoc loi nhm carboxylic, hoc c hai. Bng 3.2 gii thiu cc phn ng ny.
Cc enzym xc tc qu trnh loi amin oxy ho l cc enzym c hiu i vi cc ng phn L hoc D ca cc acid amin. Cc enzym c th l cc oxydase hiu kh (flavoprotein) hoc cc oxydase k kh, hoc cc dehydrogenase. C trng hp, trc ht nc c loi i (chuyn ho serin):
CH3 C NH COOH CH3 C O COOH Acid pyruvic + NH3
CH2OH H C NH2 COOH Serin
H2O
CH2 C NH2 COOH
+ H2O
Khi phn gii cc acid amin thm, vic ct m vng thm xy ra nh phn ng oxy ho trc tip s dng oxy, v d: phn gii tryptophan:
CH2 CH COOH NH2 N H Cat echin OH
OH
COOH COOH
Succinat t + Acet at
Acid cys, cys-muconic
Mt s vi khun (nh E. coli) c kh nng chuyn ho tryptophan thnh 2 cht c mi hi l indol v skatol. Skatol l cht to ra mi thi ca phn. S phn ng ca qu trnh ny nh sau:
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CH2CH(NH2)COOH + N H T rypt ophan H2O N H Indol + CH3COCOOH + NH3
CH2CH(NH2)COOH
CH2COOH
CH3
N H T ryptophan
N H Acid indolacetic
N H Skatol
Bn thn indol c mi hi, nhng nu cho mt cht indol vo este ca mt s hoa th li to nn mi thm c bit (c s dng trong cng nghip sn xut m phm, sn xut nc hoa), cn skatol l thnh phn chnh mi thi phn. Ngoi protein, VSV cn c kh nng phn gii cc hp cht hu c cha nit khc (nh cc base nit, ure, acid uric, kitin,...). Cc base purin, pyrimidin to ra trong qu trnh thu phn acid nucleic. Nhiu VSV c kh nng sinh tng hp ra cc enzym xc tc phn gii cc base nit ny to ra CO2, NH3 v mt s acid hu c: acid formic, acid lactic, acid acetic,.... Cc acid ny li c phn gii tip.
O HN O N H Acid uric N C O + 4H2 O N 2 (NH2)2CO + HOOC-CHOH-COOH Ure Acid tartric
Ure cng c mt s VSV phn hu (nm 1862, ln u tin Pasteur pht hin ra vi khun phn gii ure) trong c c vi khun, x khun v nm (Micrococcus ureae, Proteus vulgaris,...) do chng to ra c urease, enzym xc tc phn ng sau:
(NH4)2CO3 t bn vng nn b phn gii tip:
Kitin cng c mt s loi VSV phn gii mnh (Bacteriun chitinovorum):
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9. S PHN GII LIPID V CC ACID BO

Lipid (este ca glycerin v acid bo) v cc cht sp (este ca ru cao phn t v acid bo) l ngun dinh dng carbon v nng lng cho nhiu loi VSV. So vi cc c cht khc th cc cht ny c ng ho vi tc chm. Lipid, sp v cc acid bo cha trong xc ng vt, thc vt b vi lp vo trong t sau mt thi gian s b phn gii v ng ho ht nh hot ng ca nhiu nhm VSV khc nhau (vi khun, x khun v nm mc). Cc cht ph bt (bn cht l cc lipid) c s dng trong cng nghip, nn ngoi tc dng ph bt cng c th c cc VSV ng ho. Bc u tin ca qu trnh ng ho lipid v sp l vic thu phn cc cht ny c phn gii thnh acid bo v glycerin (hoc ru) nh lipase ngoi bo hoc ni bo. Cng c nhng nhm vi khun c kh nng to ra phospholipase xc tc thu phn cc phospholipid.
O H3C ( CH2)n C O CH2 O H3C ( CH2)n C O CH2 O H3C ( CH2)n C O CH2 Lipid
Glycerin sau khi c phosphoryl ho s c chuyn ho tip trong EMP to ra ATP v pyruvat. Acid bo c thu phn qua con ng oxy ho to ra acetylCoA (chuyn ho 2 carbon t mch), v c chuyn ho tip trong chu trnh Krebs v Glyoxylat.
CH2OH Lipase CHOH + 3CH3 ( CH2)n COOH 3 H2O CH2OH Glycerin Ac id b o
CU HI LNG GI
1. Trnh by con ng ng phn EMP v HMP, ngha ca cc con ng phn gii ng ny i vi VSV. 2. Trnh by s khc bit gia cc con ng ng phn EMP, HMP v ED, s phn gii cc loi ng khng phi l glucose. 3. Trnh by chu trnh Krebs (chu trnh tricarboxylic acid) v chui h hp. 4. Trnh by cc kiu h hp k kh, cc qu trnh ln men k kh, v trnh by s phn gii protein, lipid v cc acid bo.
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Chng 4
SINH TRNG V PHT TRIN CA VI SINH VT
Sinh trng v pht trin l thuc tnh c bn ca th gii hu sinh bao gm c cc VSV. Sinh trng l s gia tng kch thc v khi lng t bo, cn pht trin (hoc sinh sn) l s tng trng s lng t bo. Tuy nhin, phn bit r hai qu trnh nu mt cch rch ri trong a s trng hp l rt kh khn, do trong gio trnh ny nu khng xc nh r rng th khi cp n sinh trng l bao gm c pht trin v ngc li. Ngoi cc cht dinh dng cn cn c cc iu kin thch hp VSV mi pht trin c, tuy nhin, nhiu khi chng ta li cn phi s dng cc phng php tiu dit ht VSV hin c.
1. IU KIN CHO SINH TRNG VI SINH VT

Ngoi cc cht dinh dng cn c cc iu kin ngoi cnh l ho ch yu cn thit cho sinh trng ca VSV.
1.1. m
a s VSV a nc, cn nc dng t do, d hp th. Ch c mt s x khun thuc nhm a kh xerophylic s dng c nc gn trn ht t. Thiu nc xy ra hin tng loi nc khi t bo v c th dn n VSV b cht. Chu c iu kin kh l cc VSV chi Mycobacterium, v d: M. tuberculosis.
1.2. Nhit mi trng

VSV c th tn ti trong di nhit rng 2 100OC, tuy nhin, c 3 mc nhit VSV sinh trng c: nhit ti thiu sinh trng c (minimum), nhit sinh trng ti thch (optimum) v nhit cc i c th sinh trng c (maximum). Nhit khng nhng nh hng n tc sinh trng m cn nh hng n kiu sinh sn, hnh thi, trao i cht v nhu cu dinh dng ca VSV. Ph thuc vo nhit sinh trng VSV c chia thnh 3 nhm (bng 4.1).
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Cc loi Bacillus trong t c nhit sinh trng rng: 15 40OC. E. coli sinh trng c trong khong nhit 10 47OC. N. gonorrhae pht trin c nhit 36 40OC. c bit di y Thi Bnh Dng c loi vi khun a nhit pht trin thch hp nhit c th ln n hng trm bch phn (250 300OC). Ngoi cc nhm trn cn c nhm VSV chu nhit (thermotolerant), sinh trng nhit trung bnh, nhng c th chu c nhit cao hn: v d: Methylococcus capsulatas sinh trng tt 37OC, nhng cn ko di sinh trng c 55OC. nhit thp VSV khng pht trin c do thay i cu hnh lp th ca cc permease, lm bt hot cc vn chuyn xuyn mng. nhit cao VSV b cht do bin tnh protein, cc enzym, protein mng, k c ARN cng b bin tnh.
1.3. p sut, thm p, p sut thu tnh

Mng t bo cht ca VSV l mng bn thm, do p sut thm thu v p sut thu tnh cng nh hng n sinh trng ca VSV. Trong mi trng u trng, t bo mt kh nng rt nc v cc cht dinh dng ho tan xung quanh, t bo b kh sinh l, co nguyn sinh cht v c th b cht nu ko di. Nhng khi trong dung dch nhc trng, t bo VSV b nc xm nhp, nu thnh khng chc s b v. Dung dch mui 10 15%, ng 50 80% lm co sinh cht ca VSV. a s VSV pht trin tt trong mi trng c nng mui t hn 2%, nng mui cao thng c hi. C VSV a mui (halophylic) pht trin c vi nng mui n 30%, VSV a ng (saccharophylic) pht trin c vi nng ng cao, hoc a thm p (osmophylic). nhit thng, p sut cao c th lm gim ng lc ca VSV, nhng khng lm VSV cht. Nhiu vi khun di y bin chu c p sut 200 300atm, c gi l cc vi khun a p (barophylic).
1.4. Kh quyn
Trong iu kin t nhin, VSV cn mt s cht kh nh O2, CO2, N2 v CH4. Khi nui cy, c cht kh c s dng trong trao i cht ca t bo, nhng c khi li phi loi b khi mi trng do c tnh. CO2 c cc VSV s dng trong mt s phn ng. Oxy cn thit cho cc VSV hiu kh, nhng li l c i vi cc VSV k kh. Ph thuc vo nhu cu oxy ngi ta chia VSV ra thnh cc nhm sau: Hiu kh bt buc: S hin din ca oxy phn t (O2) l ti cn thit cho sinh trng ca VSV c cha enzym SOD (superoxid dismustase) v peroxydase. a s vi nm v s ng vi khun thuc nhm ny. Hiu kh khng bt buc: Sinh trng c c trong iu kin c oxy, cng nh khng c oxy, c cha SOD v peroxydase. a s nm men v nhiu vi khun thuc nhm ny: S. cerevisiae, E. coli, P. vulgaris,... Vi hiu kh: Cc VSV thuc nhm ny ch c th pht trin c trong iu kin p sut oxy rt thp (0,01 0,03ba). VSV c chui h hp oxy phn t, tip nhn hydro, bao gm Vibrio cholera, Bacteriodes spp,...
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K kh chu dng: K kh, nhng tn ti c khi c oxy, khng c chui h hp, c SOD, peroxydase, nhng thiu enzym hydrogenperoxydase, bao gm Streptococcus lactis, Lactobacillus lactis, Leuconostoc mesenteroides,... K kh: S c mt ca oxy phn t l c hi. Trong t bo khng c SOD, cytochromoxydase, phn ln khng c hydrogenperoxydase. l nhiu loi trong cc chi Clostridium, Fusobacterium, Bifidobacterium, Eubacterium, Peptococcus, Butyrivibrio, Desulfovibrio, Veillonela,...
1.5. pH mi trng
Mi loi VSV c mt pH ti thch cho sinh trng. sinh trng c trong mi trng acid hoc kim, VSV phi duy tr c pH ni bo l 7,5, bt k pH mi trng ngoi bo l bao nhiu. T bo sng c kh nng duy tr pH ni bo n nh bng cch y ra khi hoc hp th ion H+ vo t bo. a s vi khun sinh trng tt nht pH 6 8. Mt s loi sinh trng pH 9 11 (Vibrio cholerae, Bacillus sp.), vi khun lactic trong da mui chu c pH 3 4. Nm mc, nm men c khong pH sinh trng rng hn vi khun, nhng pH ti thch l 5 6. Khi VSV sinh trng mnh, pH mi trng s b thay i. S thay i pH qu ln c th s c tc ng km hm sinh trng ca VSV, v vy cn b sung cht c tnh m vo mi trng ngn cn s bin i ca pH. Ngoi cc nhn t nu vn cn nhiu tc nhn ngoi cnh nh hng n sinh trng, pht trin ca VSV, nhng do vt qu khun kh ca gio trnh nn khng th cp c, l: sc cng b mt, m thanh, nh sng, th oxy ho kh,... cng ng tham kho thm. 2. SINH SN CA VI KHUN
a s vi khun sinh sn bng con ng v tnh, ph bin l phn i theo chiu ngang mi t bo phn thnh 2 t bo con c kch thc tng ng nhau. Trc khi phn chia, ni cht t bo c tng ln gp i v th nhn c sao chp. Khi t bo m tng trng, mng t bo cng tng theo v th nhn tch bit ra thnh hai. Phn bo din ra vng gia hai th nhn khi mng t bo cht gn phn gia ca t bo tng trng tht vo bn trong, cng lc thnh t bo mi sinh cng pht trin tht vo bn trong to thnh vch ngang gia hai t bo con. T bo con c th tch ring hon ton hay vn gn vi t bo m to thnh i, nhm hay thnh chui. Mt s vi khun, v d nh Rhodopseudomonas acidophia, Mycobacterium tuberculosis sinh sn bng ny chi ging nh nm men.
3. SINH TRNG V PHT TRIN CA QUN TH VI KHUN

Sinh trng v pht trin ca qun th vi khun c cc nh khoa hc nghin cu su sc, v mt s m hnh ton hc cng c thit lp. Nhng m hnh y cng c th p dng c cho cc VSV n bo khc. Nghin cu sinh trng mt t bo vi khun nh b l rt kh khn, nn cn ch l khi ni v sinh trng v pht trin ca vi khun l cp n sinh trng v pht trin ca lng ln t bo ca cng mt loi.
3.1. Th h sinh trng ca vi sinh vt

Khi cy vi khun vo mi trng dinh dng mi, cc cht dinh dng u d tha th sau thi gian thch nghi, VSV chuyn sang thi k pht trin rt mnh m, tc sinh trng ring ca VSV khng i, khng ph thuc vo nng cc cht dinh dng. Lc ny ta quan tm n khong thi gian VSV tng sinh khi (s t bo) ln gp i. y chnh l thi gian th h (tg), hay thi gian bi i (td), c trng cho VSV trong iu kin nht nh. Thi gian th h c xc nh bng cch sau: Ti thi im t0 = 0, sinh khi VSV l x0, Sau 1 th h, t1, sinh khi s l x1 = 2x0. (3.1) Sau 2 th h, t2, ta c ... x2 = 2x1 = 22x0. (3.2)
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V sau n th h, tn, ta c xn = 2n x0 . (Vi x l sinh khi kh hay s t bo VSV). Ly logarit 2 v i vi (3.3): lgxn = nlg2 + lgx0 M n = tn/tg, vi t = tn l thi gian nui cy. Thay n vo (3.4) ta c: tg = tx lg2 / (lg xn lg x0)
(3.3)
(3.4) (3.5)
V thi gian th h (tg) thu c y l gi tr trung bnh. Mi loi VSV c thi gian th h ring, ngay mi loi thi gian th h ny cng khc nhau, nu iu kin nui cy khc nhau. V d, thi gian th h ca E. coli trong mi trng giu dinh dng l 12,5 pht, ca Mycobacterium tuberculosis l 13 15 gi. V d: Nu cy 103 t bo E. coli vo mi trng dinh dng, sau thi gian nui cy 5 gi t c 108 t bo. Tnh thi gian th h trung bnh ca qun th vi khun ang kho st ? Thay s liu vo (3.5) ta c:
Nh vy trung bnh c sau 18,06 pht, s lng t bo (hay sinh khi kh) ca qun th E. coli ang kho st li c nhn ln gp i.
3.2. Cc pha sinh trng, pht trin ca vi khun

Nui cy vi khun tnh, hay c lc trn my lc sinh hc hay trong cc bnh ln men (hay bnh phn ng sinh hc) trong mi trng dch th hay cn c gi l ln men chm l cc h thng m bo sinh trng, pht trin ca VSV c nghin cu su sc nht v c nhng c trng c th ring. Thng thng cc VSV pht trin theo 4 pha c th, tuy nhin, c nghin cu cng b m hnh sinh trng, pht trin ca VSV vi 5 pha c th, cng nh c quan nim cho rng VSV sinh trng v pht trin vi 2 pha c trng. Tuy nhin y chng ta ch cp sinh trng, pht trin ph bin nht ca VSV vi 4 pha c th m thi. Ti thi im t = 0 gi, cy ging VSV vo mi trng dinh dng ri tin hnh nui cy trong iu kin sinh l ti u. Trong qu trnh nui cy thnh phn ca h sinh khi VSV, nng cc sn phm trao i cht bin i khng ngng. V sinh trng, pht trin VSV, phn bit 4 pha c th: pha lag (pha tim tng), pha log (cn gi l pha lu tha), pha dng v pha suy tn. 3.2.1. Pha lag (pha thch ng) Khi cy cc t bo VSV t mi trng ny sang mi trng khc trong khong thi gian u, s t bo khng thay i (khng tng), nhng thnh phn t bo thay i mnh m. y l pha pht trin tim tng VSV thch nghi vi mi trng mi. Cc h thng chuyn ho ca t bo cm ng vi cc cht dinh dng mi, thi v nhn cc cht b tr (Cofactor) thit yu qua khuch tn v cc enzym trao i cht dn thch nghi vi cc iu kin mi ca mi trng. Trong vic hnh thnh pha lag ny, trng thi sinh l ca ging c ngha quyt nh. Nu ging l cc VSV th sinh dng (vegetative) th VSV hu nh pht trin lun, thi gian pha lag ngn. Nu ging l cc t bo dng khng pht trin (bo t, nh bo t, VSV t mi trng ngho,...) th thi gian pha lag cn di thch nghi. 3.2.2. Pha log (pha lu tha) Cui pha lag, cc t bo thch nghi vi iu kin sng mi v bt u pht trin mnh m qun th VSV chuyn dn sang pha pht trin mi l pha log, pha pht trin nng ng nht. Nu ta gi x l nng sinh khi kh ca VSV trong dch ln men th dx/dt l tc tng trng sinh khi trong 1 n v thi gian. Gi l tc tng trng sinh khi ring ca VSV, ta c:
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1 dx x x dt
(3.6)
Gi s = khng i (const), li c:
dx = dt x dx x = x0
x
dt
0
(3.7)
ln
x = t ln x = t + ln x 0 x0
(3.8) (3.9)
Vy di dng hm logarit, sinh trng, pht trin ca VSV c dng hm tuyn tnh, do , pha ny c tn gi l pha log. Tuy nhin, hm lu tha cng c s dng biu din qu trnh ny, v vy cng cn gi l pha lu tha. Khi dinh dng cn dn, nghin cu v sinh trng, pht trin ca VSV, Monod thy rng, tc sinh trng ring ph thuc vo nng cht c ch cht bt u thiu ht (S), tc sinh trng ring cc i (m) v h s c hiu c cht Ks: (3.10) Gi tr Ks chnh l nng c cht m ti tc sinh trng ring chnh bng 1/2 tc sinh trng ring cc i ( = 0,5 x m). Tuy nhin, nu cng mt lc c nhiu cht dinh dng ang cng c ch th phi m rng cng thc trn:
(3.11) Nu tt c cc cht dinh dng u d tha, tc l = m, VSV pht trin trong pha log. Nu 1 cht dinh dng b cn kit, do cn cc cht khc m xut hin 1 pha log khc vi tc sinh trng ring khc (2). Trong thc t cc gi tr ca KS rt b. V d: E. coli c KS, glu = 1,0mg/l, KS,trypt = 1,1mg/l. Cc gi tr KS rt b ny chnh l li gii thch cho qu trnh ln men gin on m ti thi im pha log h thng khng t n cc gi tr KS rt b ny. Tuy nhin, v lng sinh khi ln v ngy cng tng ln m trong thi gian tng i ngn do cn kit cc cht dinh dng bt u giai on khi im ca pha dng. Tuy nhin, m hnh Monod khng ng dng c trong nhng trng hp m VSV pht trin rt nhanh: mc d c cht dinh dng trong mi trng, nhng nng ca chng trong min ln cn t bo tr thnh c ch. * Hin tng sinh trng kp: Trng hp mi trng dinh dng phc hp thng nhn thy 2 pha log, gia chng c 1 pha lag phn cch. Hin tng ny gi l hin tng sinh trng kp, do trong mi trng c 2 loi hydratcarbon: trc tin VSV phn gii 1 cht (thng l glucose). S c mt glucose ngn tr s phn gii cc cht khc. Ch sau khi ng ho ht cht dinh dng u, trong pha lag th hai, cc enzym cm ng mi c to ra s tip tc ng ho cht cn li. 3.2.3. Pha dng hay pha n nh
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Sau khi ng ho cc cht dinh dng hay sau khi tch lu cc sn phm trao i cht, sinh trng ca VSV gim xung hay hon ton ngng li. Trong pha n nh ny, sinh khi c th cn tng chm hoc khng i. Do s t phn ca mt s t bo, mt s cht dinh dng mi (hydratcarbon, protein) c gii phng, iu ny a li s pht trin chm ca mt s t bo sng st. Trong pha dng ny, rt nhiu sn phm trao i cht c gi tr (khng sinh, vitamin,...) c to thnh. 3.2.4. Pha suy tn c trng ca pha ny l cc cht dinh dng b cn kit, nng lng ca t bo gim n ti thiu. T bo cht i theo quy lut ging nh khi tng trng trong chu k sinh trng (tuy nhin vi khuynh hng ngc li) (hnh 4.1).
c trng ca pha dng hay pha suy tn ph thuc vo tng loi VSV v vo phng php nui cy c ng dng. T quan im cng ngh, ngi ta kt thc qu trnh nui cy vo cui pha log, trong pha dng hay pha suy tn ph thuc vo vic sinh tng hp hot cht b nh ch hay tr nn khng cn kinh t na pha no trong chu k sinh trng ca VSV.
4. CC SN PHM TRAO I CHT CA VI SINH VT

Ngoi cc sn phm chuyn ho nu trn cng nh cc cht l thnh t to thnh cu trc t bo, VSV cn to ra hng lot cc sn phm trao i cht khc. Cc cht do vi khun ni ring v cc VSV ni chung to ra c th l c hi i vi con ngi, nhng cng c th l cc sn phm hu ch c s dng trong y dc hc cng nh trong cc ngnh kinh t k thut khc.
4.1. c t
c t l sn phm c to ra trong qu trnh sinh trng, pht trin ca nhiu vi khun gy bnh. C hai loi c t: Ngoi c t l cht c c vi khun, vi nm tit ra ngoi c th, thng l c t ca trc khun G+ hay ca nm gy bnh. Ngoi c t c c tnh rt mnh, v d: ch cn 0,02mg ngoi c t bch hu, hoc 0,0006mg ngoi c t un vn l c th gy cht ngi. c t ny l protein tan c trong nc.
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Ni c t l c cht ca trc khun G, ca cc vi khun ng rut. c tnh khng mnh bng ngoi c t, v d: ni c t thng hn phi cn ti 400mg mi gy cht ngi. Bn cht ni c t l phc hp glucidlipidprotein. Ni c t nm bn trong t bo v ch c gii phng ra ngoi khi cc t bo b ph v.
4.2. Cht gy st
Mt s vi khun c kh nng to ra hp cht gy st (pyrogen), khi tim cho ngi hay sc vt gy nn phn ng st. Cht gy st khng b nhit ph hu nn sy, hp khng ph hu c cht gy st. Mun loi b c cht gy st phi lc qua phu lc thu tinh G5 hay mng lc aming. Nc dng pha thuc tim nht thit khng c php cha cht gy st.
4.3. Cc vitamin
Nhiu loi VSV c kh nng sinh tng hp c vitamin (nhm B, C, D,...). Ergocalciferol (Vitamin D) c nhiu trong nm men (Sac. carlsbergensis), nm mc Asp. niger, Penicillium,..., tuy nhin vi khun t hn. Thiamin (Vitamin B1) c trong Pseudomonas fluorescens, Torulopsis utilis,... Pyridoxin (Vitamin B6) c trong Aerobacter aerogenes, C. butyricum,... Tuy nhin, ch c hai vitamin c sn xut trn quy m cng nghip bng ln men VSV, l vitamin B2 (riboflavin) nh nm Eremothecium ashbyii v Ashbya gossypii, trong hiu sut ca Ashbya gossypii l cao hn. Vitamin B12 c sn xut nh ln men Pseudomonas denitrificans, Propionibacterium shermanii,...
4.4. Cc khng sinh

4.4.1. nh ngha khng sinh Khng sinh l lp hot cht hu ch c tc dng sinh hc rt mnh (nu khng ni l mnh nht) m VSV to ra c, do c nhiu nh khoa hc quan tm nghin cu. C nhiu cch nh ngha v khng sinh: nh ngha hp, nh ngha rng,... song nh ngha cht ch theo Waksmann (1942) c ngha kinh in: Khng sinh l nhng sn phm trao i cht t nhin c cc VSV to ra, c tc dng c ch pht trin hoc tiu dit chn lc i vi cc VSV khc. Tuy nhin nh ngha ny kh hn ch, v nh vy, nhm cc khng sinh thc vt, cng nh cc khng sinh bn tng hp, cc khng sinh c tng hp ton phn v cc hp cht ho hc c tc dng tiu dit chn lc cc VSV nhim sinh khc u khng c cp n. Ngy nay, mt cch cp thit, thut ng khng sinh theo ngha rng c th s dng (tuy nhin cc nh khoa hc vn cha ht tranh lun) i din cho lp tt c cc hp cht ngun gc t nhin hoc tng hp c tc dng c ch hoc tiu dit chn lc i vi cc VSV nhim sinh (cng nh c vi t bo ung th) nng thp, m khng c tc dng hoc tc dng yu ln ngi, ng vt, hoc thc vt bng con ng cung cp chung. Khng sinh l sn phm trao i cht th cp ch c sinh tng hp mnh m giai on pht trin sau (pha logarit mun, pha dng, thm ch pha suy tn) ca sinh trng VSV. 4.4.2. Mt s khng sinh c quan tm Vi khun, nm, thc vt, v.v,... c th sinh tng hp c khng sinh, tuy nhin y chng ta ch cp n cc khng sinh c sn xut nh vi sinh tng hp m thi (bng 4.2).
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Vi: F nm; Myco. Mycobacterium.
4.4.3. C ch tc dng ca khng sinh Mi khng sinh u c ch tc dng nht nh trong cc t bo VSV mn cm, tuy nhin c th khi qut thnh 6 nhm ch tc dng chnh (hnh 4.2): 1. Tng hp thnh t bo: lactam (penicilin), vancomycin, bacitracin. 2. Mng t bo cht: amphotericin, polyen, valinomycin. 3. ADN: actinomycin, antracyclin, anzamycin (rifamicin). 4. Tng hp protein: ribosome (aminoglycosid), Lin kt tARN (macrolid), chlorocid, ko di mch polypeptid (tetracyclin, acid fuzidic). 5. Trao i cht h hp: antimycin, oligomycin.
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6. Trao i cht folat: sulfamid,...
4.4.4. C ch khng thuc khng sinh ca vi khun 4.4.4.1. Tnh khng thuc Khi x l VSV vi cc khng sinh khc nhau, nhn thy rng: Mt s vi khun nh Pneumococcus, Gonococcus, Meningococcus, Haemophilus influenzae c nhy cm khng sinh kh ng nht. Cc vi khun khc: Staphylococcus, Enterococcus, E. coli, Klebsiella, Pseudomonas pyocianae,... li c mn cm rt thay i, do kho st nhy khng sinh rt kh khn. Tuy nhin vic s dng khng sinh cha hp l, hay ni ng l s dng sai, trn lan khng sinh trong iu tr cng nh trong chn nui xut hin ngy cng nhiu cc VSV khng khng sinh (nhn thuc) lm cho khng sinh khng cn hiu nghim na. S xut hin ca VSV khng khng sinh v cc cht ho tr liu khc l vn thi s ca Y sinh Dc hc hin i. nh ngha: Khng thuc l hin tng VSV mt i tnh nhy cm ban u ca n trong mt thi gian hay vnh vin vi tc dng ca khng sinh hay ho tr liu. C 2 kiu khng thuc: Khng thuc t nhin v khng thuc mi nhn (Acquired Resistance). a) Khng thuc t nhin Khng thuc t nhin l c trng ca tng ni VSV nht nh i vi mt s khng sinh nht nh no . Tnh cht ny c sn trc khi s dng cc khng sinh . iu ny lin quan n ph tc dng ca khng sinh, c th c trng cho tng loi (Proteus khng cc tetracyclin), cho tng nhm (trc khun G khng penicillin G), tng sn phm (Streptococcus nhm D khng lincomycin). V mt sinh ho, c 2 c ch quan trng: tnh thm ca t bo v s thiu vng phn t ch. b) Khng thuc mi nhn Xut hin trong chn lc t nhin cc chng khng ca qun th VSV nhy cm khi s dng khng sinh. Mt VSV tr thnh khng thuc khi c th pht trin c vi hm lng cao ng k ca khng sinh y so vi qun th VSV m n c ngun gc. khng sinh hc (biological resistance) l hin tng c nhng c th ca mt loi do thu c nhng c tnh di truyn m gim thiu hoc mt hn tnh nhy cm thuc so vi nhng c th cng loi. Khi mc khng tng ln, chng thot khi s iu tr khng th kim sot bnh bng khng sinh c hay bnh khng cha c ta ni n khng iu tr (therapeutical resistance) hay khng thuc lm sng. Nhng c th khng sinh hc khng nht thit phi l khng iu tr.
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4.4.4.2. C ch di truyn hc tnh khng thuc Trn phng din di truyn hc, khng thuc mi nhn c th l do s thay i gen nhim sc th (t bin nhim sc th), c th do tip nhn gen plasmid ( khng plasmid hay ngoi nhim sc th). a) Khng thuc do t bin nhim sc th Di tc dng ca khng sinh vic tiu dit cc vi khun mn cm to ra s tuyn chn ngu nhin cc t bin mi xut hin v chng c pht trin nhn ln. t bin kiu ny t n nh v t di truyn. Kiu t bin ny chu nh hng ca tng nhm khng sinh: c dng xut hin rt mnh m (mt bc kiu streptomycin), hoc t t (nhiu bc kiu penicillin). Kiu khng thuc ny chim khong 10% tng s VSV khng thuc, xut hin ch yu i vi cc khng sinh nh: lactam, aminosid, cloramphenicol, erythromycin, rifamicin,... v cc khng sinh tc dng vi cc Mycobacteria (hnh 4.3).
b) Khng thuc plasmid Kiu khng thuc ny rt ph bin, chim 90% s VSV khng thuc. Khng thuc ngoi nhim sc th l hin tng a khng thuc do nhn t R (Resitance factor plasmid). Nhn t R chnh l cc plasmid cha cc gen iu khin tnh khng thuc. Cc gen ny thng qua cc c ch di truyn ti np (transduction), bin np (transformation), tip hp (conjunction) chuyn ti thng tin di truyn khng thuc t t bo ny sang t bo khc, loi ny sang loi khc. Cc nhn t R plasmid l nhng n v di truyn ngoi nhim sc th, bao gm nhng phn t ADN dng vng khp kn c ln khc nhau vi phn t lng t 1 260 MDa, tng ng khong 3 500 gen v c kh nng t sao chp, tn ti trong cc t bo VSV. Hin tng khng cho l hin tng vi khun khi khng mt khng sinh th ng thi khng lun mt s khng sinh khc c cu trc tng t. V d: cc chng Staphylococcus khng penicilin G v cc lactam khc nh ampicilin, cephalosporin. 4.4.4.3. C ch sinh ho khng thuc mi nhn C 4 kiu chnh: a) Thay i tnh thm thnh t bo t c iu ny c th thay i enzym chu trch nhim tnh thm, v vy khng sinh khng thm qua c thnh t bo (khng lactam, hay tetracyclin) khng pht huy tc dng c, hay c th thay i khng sinh bng enzym (aminosid). b) V hiu ho cc khng sinh bng enzym lactam: Cc lactamase ph v vng lactam m v hiu ho khng sinh.
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H R CO N N O
CH3 CH3 COOH
V tr tc dng ca lactamase
H R CO N N O
S R' COOH
V tr tc dng ca lactamase Cc lactamase khc nhau v h s enzym v tnh c hiu c cht v im ng in. C th chia cc lactamase thnh: penicillinase, cephalosporinase v penicillinase ph rng. Cc aminosid b thay i khng qua c thnh t bo, cng nh b mt tc dng do cc enzym l adenylase v phosphorylase tc dng ln cc v tr nhm OH v acetylase acetyl ho cc nhm RNH2 (chng c tnh c hiu c cht, v tr,...).
Cloramphenicol b mt hot tnh do enzym acetyltransferase plasmid
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Mt hot cloramphenicol. c) Thay i phn t ch Phn t ch, ni tc dng ca khng sinh b thay i, khng sinh khng cn ni lin kt, v d: thay i protein ribosom lin kt vi streptomycin hay thay i ARNpolymerace lin kt vi rifamicin. d) Hot ho con ng trao i cht thay th khc m hot cht khng tc dng. 4.4.4.4. Nguyn tc s dng khng sinh Nh vy vic s dng khng sinh cha hp l trong iu tr lm xut hin ngy cng nhiu VSV khng thuc. khc phc hin tng , cn thc hin mt s nguyn tc sau: Phn lp chng VSV gy bnh v th nhy ca chng vi cc khng sinh (lp khng sinh ) bng phng php khoanh giy lc,... Chn khng sinh c hot tnh mnh nht. Quyt nh liu dng, cch a khng sinh vo c th, v iu tr. Phi hp khng sinh vi ch phm khc (corticoid, enzym) lm tng tc dng, gim phn ng ph.
5. CC TC NHN ST KHUN
Cc tc nhn st trng c hiu nh l cc tc nhn tiu dit cc t bo, bo t, nh bo t VSV,... m khng c tnh chn lc. Hng lot cc nhn t vt l v ho hc c xp vo nhm tc nhn ny.
5.1. Cc tc nhn vt l
Cc tc nhn vt l bao gm nhit cao: nhit m hay nhit kh. Nhit m tiu dit VSV hiu qu hn nhit kh v nhit m gy ng vn v bin tnh enzym, cn nhit kh lm oxy ho cc thnh phn hu c ca t bo. Bn cnh nhit cc bc x cng l cc tc nhn vt l dit khun hiu qu. Tia ion ho bao gm tia gama v tia X. Cn nh sng UV tuy khng c kh nng ion ho, nhng do c kh nng m xuyn n 3m, v c ADN hp th bc sng 260nm (thng s dng 254nm) nn c tc dng dit khun mnh, c s dng kh trng b mt, hay kh trng cc dung dch trong sut vi cc thit b c dng.
5.2. Cc tc nhn ho hc
Nhiu ho cht l tc nhn st trng tt, trong s c th lit k di y: Phenol v cc cht cng h, Ethanol: Ethanol 70% c tc dng st trng mnh nht, c dng st trng bn ngoi. Halogen v cc cht cha clo hot ng bao gm: Iod v cc hp cht cha iod; clo v cc hp cht cha clo hot ng; v d: cloramin B, T. Cc cht cha oxy hot ng nh: H2O2, KMnO4.
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Cc kim loi nng: thu ngn clorid (HgCl2), cc hp cht hu c cha thu ngn c hot tnh khng khun cao v c tnh thp nh: mercurocrom, timerosel, nitromersol (metaphel) c dng x l vt thng da; cc phc cht hu c cha bc, hay AgNO3 1%; cc hp cht km nh km sulfat. Cc cht ty ra (detergent), x phng. Formaldehyd: formaldehyd l cht kh kh bn vng nhit cao, rt c, kch thch mnh cc mng nhy. Dung dch nc 37 40% formaldehyd cn c gi l formalin c dng kh trng dng c. Dng kh dng ty trng khng kh. Dung dch 10% trong cn c dng v trng phng th nghim rt c hiu qu.
CU HI LNG GI
1. Trnh by cc iu kin cn thit cho sinh trng ca VSV, trnh by cc giai on pht trin ca VSV trong mi trng lng. 2. Trnh by tng qut cc sn phm ca VSV, c ch tc dng ca khng sinh i vi vi khun. 3. Trnh by tnh khng thuc ca VSV. 4. Trnh by cc tc nhn st khun v ng dng.
Chng 5
DI TRUYN VI SINH VT
1. I CNG
Cc VSV cng u ging t tin mnh hu ht cc c im. Di truyn l vic duy tr cc c im ca t tin cho i sau ca th gii hu sinh. n v ca di truyn l gen. Gen l mt on ADN m nhim vic m ho mt tnh trng di truyn, cc c im ca gen c di truyn li cho i sau qua sao chp. Mt khc gen m nhn chc nng nht nh trong qu trnh truyn thng tin di truyn, m ho cho mt chui polypeptid m c c m qua ARN (mARN, tARN, v cc loi ARN khc), c chc nng iu chnh hoc ng vai tr iu khin s biu hin hot ng ca tng gen (genom). Mt s virus c vt liu di truyn l ARN (virus cm, virus di, HIV,...) th gen l mt on ARN m ho cho mt protein xc nh c c m qua b my phin dch ca t bo ch. Phn ln gen nm trong nhn t bo, phn nh nm trong yu t di truyn ngoi nhim sc th chng hn plasmid (plasmid F, plasmid R), v cc yu t di truyn di ng (yu t IS v transposon).
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Theo quan nim hin hnh dng thng tin di truyn t nhim sc th n t bo cht mi VSV din ra nh sau:
Bn thn cht di truyn (ADN hoc ARN) c kh nng t nhn ln, qu trnh ny c gi l sao chp (1). Mt khc ADN c dng lm khun tng hp ARN (mARN, tARN v rARN) trong qu trnh phin m (2a). Mt s virus (chng hn Retrovirus) m c cht di truyn l ARN, c th gn genom ca chng vo nhim sc th ca t bo ch phi tng hp dng ADN trung gian t si khun ARN. Qu trnh ny c gi l phin m ngc (2b). Cui cng, dch m hay sinh tng hp protein (3a) din ra trn phc hp bao gm cc si mARN, ribosom (c cha cc rARN) v t ARN (mang acid amin). Tuy nhin, liu qu trnh 3b c tn ti trong t nhin hay khng? cn ang trong qu trnh ch i cc nh khoa hc tr li trong tng lai. Ta phn bit genotyp (kiu gen b my di truyn ca mt t bo) v phenotyp (kiu hnh biu hin bn ngoi ca b my trn) ca VSV. Genotyp l tng tnh trng di truyn trong mt c th, nhng cha chc c biu hin ht, m trong mi tng quan vi mi trng ch mt kiu hnh (phenotyp) trong c biu hin m thi. Cn phn bit genom (h gen, b gen, tng gen) ca c th sng v genotyp (kiu gen) ca c th sng ni ring.
2. SAO CHP ADN, PHIN M, DCH M
2.1. Sao chp ADN

Qu trnh sao chp c nghin cu k nht E. coli. Nh bit, h gen (genom) ca E. coli l mt ADN si kp, dng vng c mt im khi pht sao chp (oriC), l mt n v replicon. Sao chp bt u im gc oriC v tip tc cho n kt thc. Khi mt s protein nhn ra im oriC, hai si n ADN s c tch ra to thnh chc sao chp, ADN s c tng hp theo hai hng ngc nhau: trn si th nht tng hp tin theo chiu 5 3 mt cch lin tc, cn trn si th hai cng t 5 3 nhng tng hp tng on ADN theo hng li, ri gn cc on li vi nhau. Cc enzym v protein tham gia tng hp ADN gm: gyrase (topoisomerase II), enzym tho xon; helycase gn vo 2 si n, tip tc ci xon ADN; protein lin kt si n (SSB) gi cng si n; ADN polymerase, phc hp enzym c 3 thnh phn: pol I, pol II, pol III. C ba ADN polymerase u c kh nng sao chp ADN theo hng 5 3, cn c mt on 3OH gn nucleotid vo. u 3 OH ny do mt on ARN ngn (mi primer do primase to ra) cung cp. Primase gn vi polypeptid to thnh primosom, c kh nng nhn bit th t c bit trn ADN tng hp mi. ADN polymerase c c hot tnh nuclease. V chc nng ADN pol III tng hp, ADN pol I tng hp b sung v sa cha, ADN pol II h tr. Tng qut li sao chp ADN E. coli din ra nh sau: Mt s protein (proteinB) nhn ra gc oriC trn ADN v gyrase tho xon y, 2 phn t helicase gn vo 2 si n ADN tip tc ci xon, cc protein SSB gn vo si n sau helicase, trn si 3 5 primase tng hp mi primer (duy nht) v pol III gn tip vo 3'OH ca cc nucleotid tng ng, ri c th tip tc (si con c gi l si trc) hng vo ch 3. Trn si khun th hai, primase tng hp nhiu mi, pol III gn tip vo u 3'OH ca cc nucleotid v ko di cc on nucleotid n 1.000 2.000base (on Okazaki) hng ra ngoi. Khi pol III bt gp on mi ARN trc th dng li, pol I ct b mi ng thi gn b sung cc nucleotid vo, ligase hn khe h gia cc on Okazaki (cn tiu th NADH2). Nh vy, si con to thnh trn si ADN khun 5 3 c sao chp theo kiu gin on v c gi l si mun (hnh 5.1).
AND Protein ADN Khun ADN ARN Topoisomerase Primase ADN M(ARN
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Hnh 5.1. Sao chp ADN E. coli
Sinh tng hp ca 2 si con c phi hp nh to thnh mt nm trn si mun gn chc, nh vy dime ca pol III c th sao chp ng thi cng hng c hai si. Tng hp ADN phage X174 ch theo mt hng, v ch cn pol I v ligase, v cng khng cn ARN lm mi. C th tn ti trong t bo replisom ging nh ribosom cho sinh tng hp protein.
2.2. Phin m
Phin m trong t bo VSV cng din ra theo hng 5 3. E. coli enzym xc tc phin m sinh tng hp c 3 loi ARN l ARN polymerase bao gm 5 chui peptid: 2, , ', (holoenzym). 4 Chui peptid u gn cht vi nhau v c hot tnh polymerase gi l enzym ti thiu. Chui s gn lng lo vo enzym ti thiu, gip enzym ny gn chnh xc vo v tr m u (promoto) ca mi gen. Phin m ch din ra trn mt si, thm ch ch trn tng on ca ADN, mt khc, ARN polymerase khng cn mi v cng khng c hot tnh nuclease. Phin m E. coli din ra nh sau: ARN polymerase gn vo promoto trn ADN v ci xon y nh ch ch ca yu t . Phin m bt u gn ATP hoc GTP (lm nucleotid th nht) vo chui . Sau khi phin m c 12 nucleotid, tch khi phc hp v phin m tip tc. Khi phin m gn n kt thc gen, ARN polymerase s gp mt trong hai dng tn hiu kt thc: tn hiu kt thc mnh dng kp tc, hoc tn hiu kt thc yu cng c dng kp tc, nhng thiu yu t olygo U v cn yu t protein rho; rho nhn ra v gn vo on ARN si n, thu phn ATP ri di ng n bng phin m v tch mARN khi phc hp. Sau khi tng hp da trn si khun, ARN cn phi c ch tc tr thnh dng cui cng tham gia vo chu trnh sng ca t bo, l tch cc nucleotid tha, ci bin cc nucleotid v b sung cc nucleotid. Tch cc nucleotid tha: Ni chung khong 22% nucleotid ca sn phm phin m ban u b ct i. V d: s hnh thnh cc rARN 16S v 23S t cc p 16S v p 23S,... Ci bin cc nucleotid: Trong qu trnh trng thnh ca cc ARN thng xy ra ci bin, v d: thc hin gn methyl vo mt nucleotid t adenosylmethionin nh methyltransferase, hoc isomer ho uridilat thnh pseudouridilat, hay uridin thnh thiouridin,.... B sung cc nucleotid: Trnh t CCA c thm vo ARN khi phin m nh tc dng ca
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nucleotidyl transferase, v enzym ny cng c chc phn thm trnh t CCA vo tARN b loi mt CCA sau mi vng trao i cht.
2.3. Dch m
Dch m cng c nghin cu k nht E. coli vi 3 b phn chnh ribosom, mARN v tARN. Nh bit, Procaryota, ribosom thuc loi 70S c cu thnh t 2 n v nh 50S v 30S. n v 50S cha 34 protein (L1 L34) v 2 loi rARN (23S v 5S), n v 30S cha 21 protein v mt loi rARN 16S. Trn ribosom c 2 v tr gn tARN: v tr A gn aminoacyltARN v v tr P gn peptidyltARN. tARN ch cha 70 90 nucleotid, gm nhiu base ci bin (dihydropseudo thiouridin, inosin, ribothymidin,...) c cu trc l ch ba vi cung v thu 3. u 5 ca cung lun tn cng bng CCA, l v tr gn acid amin, cn 3 thu ln lt c gi l DHU (dihydrouridin), AC (anticodon) v TX. M u dch m lun l methionin, v vy t bo cn 1 loi tARN vn chuyn methionin vo u chui (tARNfMet) v 1 loi vn chuyn methionin vo gia chui (tARNmMet). Acid amin c hot ho trc khi tham gia tng hp protein:
Acid amin + ATP
aa~AMP + tARN
aa~tARN + AMP
E. coli v cc t bo procaryota, mARN l polycistron (c m cho hn 1 polypeptid) v hu nh phin m v dch m xy ra ng thi, do m hu nh khng c mARN nguyn vn. Dch m c 3 giai on: m u, ko di v kt thc. 2.3.1. M u Tham gia vo giai on ny cn c thm 3 tc nhn protein IF1, IF2, IF3. Trc tin, nh kch thch, IF3, mARN lin kt c vi ribosom 30S, ri phc fMetARNfMetIF2GTP c chuyn vo v tr P trn phn 30S ng vi codon u AUG ca methionin (vi s h tr ca IF1). Nh trnh t 3 9 nucleotid bt cp vi nhau trn u cui 5 ca mARN v u cui 3 ca rARN 16S (tn hiu ShineDalgarno, hay v tr lin kt ribosom, RBS) m codon dn u AUG c chuyn chnh xc vo v tr khi u. Lc ny, IF3 b tch ra. Khi fMetARNfMet gn chnh xc vo v tr m u th ht 50S gn vo thnh monosom 70S, ng thi GTP b IF2 thu phn, nng lng gii phng ra y c IF1 v IF2 ra, v phc khi u [70SmARNfMetARNfMet] c to thnh. 2.3.2. Ko di Giai on ny cn c s tham gia ca hai yu t protein ko di: EFT v EFG. Yu t T li c cu thnh bi Tu v Ts lin kt lng lo vi nhau. Lin kt ca acid amin th 2 (aa~tARN) tr i vo ribosom cn s kch thch ca Tu v GTP trong phc aa~tARNTuGTP. Tip theo, [aa2~tARNTuGTP] gn vo v tr A ca ribosom vi codon tng ng, v tng t
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nh bc u GTP b Tu phn gii v TuGTP b y ra. Nh Ts m Tu li c ti hot:
Nh chuyn nhm COOH ca fMet sang nhm NH2 ca aa2 bi peptidyltransferase (ribozym 23S ca ribosom 50S) m lin kt peptid th nht c to thnh. Lc ny, fMetaa2 tARN dch chuyn t A sang P v y tARNfMet trng ra ngoi. Tip tc, [aa3~tARNTuGTP] li gn vo v tr A v cc bc c th tip din cho n kt thc. 2.3.3. Kt thc Khi tng hp xong chui polypeptid ribosom s gp mt trong 3 codon kt thc hay codon v ngha UAA, UAG hay UGA. vi khun c 3 protein tham gia vo giai on ny: RF1 (UAA, UAG), RF2 (UAA, UGA), v RF3 kch thch hai protein trn. Ngoi ra, yu t tch ri RRF ca ribosom v yu t ko di EFG cng cn thit. Chui polypeptid b tch khi tARN cui cng, phn ng ny c xc tc bi peptidyl transferase, v ribosom cng b phn ly. Tuy nhin, dch m khng phi ch din ra trn mt monosom m din ra trn nhiu monosom k cnh lin kt vi nhau gi l polysom.
3. PHT SINH T BIN V CC KIU T BIN

Cng nh sinh vt bc cao, t bo VSV cng chu t bin. t bin vi khun chu tc dng trc tip ca mi trng. Ni chung, t bin (mutation) l s thay i kiu gen ca VSV, tuy nhin cng c th t c iu ny nh ti t hp (recombination). t bin c th l ngu nhin, hay nhn to (gy to). Ti t hp c th l hu tnh hay cn (gi) hu tnh (parasexual recombination) hay ti t hp di truyn.
3.1. t bin ngu nhin v t bin gy to
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3.1.1. t bin ngu nhin Trong qun th VSV lun din ra t bin ngu nhin vi tn sut khong 1010 105. Nguyn nhn c th l do tc ng ca mi trng v do chuyn ho tautomer (h bin) ca cc base khi sao chp, v d: T tn ti bnh thng trng thi keto (=oxo) s ghp i (sng i, bt cp) vi A. Nhng khi sao chp ng lc T h bin chuyn sang dng enol (hydroxy) s ghp i vi G. Hu qu l trong si ADN mi sau mt th h, mt cp GC s thay vo v tr l ra l ca cp AT. 3.1.2. t bin gy to Khi x l t bo VSV vi cc tc nhn t bin (vt l, ho hc, sinh hc hay kt hp cc tc nhn y) i a s VSV b git cht. Trong s cc VSV sng st xut hin cc c th mang t bin. t bin nh th c gi l t bin gy to. Tc nhn t bin c th l vt l, ho hc hay sinh hc. V tng th c hai dng t bin: t bin tng gen (genom) v t bin gen. t bin tng gen l lm thay i s genom ca sinh vt (v d: t n bi thnh a bi), c gi tr trong ci to ging thc vt. t bin gen l lm thay i cu trc ca gen. t bin ny c hai kiu chnh: t bin im v t bin chuyn (trt) khung. t bin im xy ra khi mt cp base b th bng mt cp base khc, v d: AT b thay bng GC, y l s chuyn dch (transition) base purin c thay th bng base purin; cn khi AT b thay bng CG, y l s o dch (transversion) base purin c thay th bng base pirimidin. Hnh 5.4 gii thiu t bin i cp (chuyn dch).
AT Tc nhn t bin (H)A* T (H)A* C AT Cp baz A di tc dng T ca tc nhn t bin bin i thnh cp G trong allen C t bin.
(H)A* C
GC
Hnh 5.4. t bin im i cp base
t bin chuyn khung (trt khung) xy ra khi mt on ADN gm mt s base, thm ch nhiu gen b loi i, b chuyn ch, hoc b cch ra do s xen vo (chn vo) ca on ADN l. Hnh 5.5 gii thiu t bin trt khung.
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C C G CT TT T C G A T G G C G A A A A G C TA t on
C C G C T T T TC G A T GGCG A A A G C TA A Ti tng Sng i sai hp ADNligase
CCGC T TTTCGAT GGCGA AGCTA Ti tng hp ADNligase Phn hu
CCGCTT T T CGAT G G C G A A A A G C TA A Thm A
TT CCG C TT CGAT G G C G A A G C TA Mt A A
Trt khung thay i m (cc acid amin thay i trong protein)

Hnh 5.5. t bin trt khung
3.2. C ch tc dng ca cc tc nhn t bin

3.2.1. Lp cht tng t base Cht tng t base l cht khng trao i (antimetabolite), t bo nhm ln lp vo ADN, thng dng gy t bin l BU (5bromouracil) v AP (2aminopurin). BU tng t nh T nn trong ADN BU chim ch T sng i vi A. Tuy nhin, do khuynh hng d tautomer ho thnh dng enol m vng sao chp tip theo BU s sng i vi G. Hu qu l t bin im i dch TA thnh CG. AP cng tc dng tng t: mc d th hin nh A, nhng thng tautomer ho thnh dng imino sng i vi C. 3.2.2. Thay i ho hc ca base Acid nitr (HNO2) kh amin ca A hoc G nhng khng lm t si, do thay th nhm NH2 bng OH, nn: A thnh hypoxantin ghp i vi C, do i AT GC t bin. G thnh xantin, vn sng i vi C nn khng gy t bin. Hydroxylamin phn ng ch yu vi C, khin base ny sng i vi A, do cng dn n t bin: CG TA. Ethyl v methylsulfonat, ethylenimin, Nnitrosoguanidin l cc tc nhn alkyl ho gy t bin mnh. V d: EMS (ethylmethansulfonat) ethyl ho ch yu N7 ca guanin. 7alkylguanin b tch khi chui li mt l hng, khi sao chp, mt base sai thng lp vo y. 3.2.3. Chn thm vo hoc loi i mt cp base Phn t acridin xen vo gia cc cp base trn chui ADN lm tng khong cch gia chng, vic ny s lm mt i mt cp base nucleotid hoc lp thm vo mt cp base v nh vy lm chuyn khung c trong tng hp protein, gy t bin. 3.2.4. nh sng t ngoi UV v cc bc x ion ho
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nh sng t ngoi UV, tia Rnghen v cc bc x ion khc c tc dng gy cht mnh v gy t bin. nh sng UV, 253 280nm, tc dng ln acid nucleic dn n dime ho thymin (TT), thymin cytosin (TC) hoc cytosin (CC) ti v tr C4 v C5. Hu qu l sao chp b sai lch v ADNpolymerase d lp mt nucleotid khng chnh xc vo v tr trn. Cu trc ca dime TC c cng thc sau:
O N O N R N R CH3 NH2 N O
nh sng t ngoi thng gy ra cc t bin i dch GC AT, trt khung, thm ch mt on. Tia Rnghen, tia v tia c s dng khi cc tc nhn t bin khc khng s dng c (v d khi t bo cht khng cho nh sng i qua). Khi s dng cc tia ion ho th xc sut to ra s t on trn c 2 si ADN s tng ln mnh m. Cc t on trn mt si s c c ch ct b nucleotid sa cha 90%. C ch sa cha ph thuc recA (sa cha hu sao chp post replication repair) cng tham gia. Nu c hai si cng b t th c th ko theo cc thay i nghim trng nh chuyn v (translocation), o cp (inversion), cng nh cc t bin nhim sc th tng t khc.
4. TI T HP DI TRUYN V S CHUYN TNH TRNG
4.1. Ti t hp di truyn
t bo nhn nguyn thu cng xy ra ti t hp, tuy nhin, y ti t hp c khc bit so vi t bo nhn tht. Hp t khng phi l sn phm kt hp ca hai t bo, thng ch mt phn ADN ca t bo cho c chuyn sang cho t bo nhn, do , ch xut hin hp t mt phn. ADN ca t bo nhn v on ADN ca t bo cho sng i v trao i on, v khi phn bo tip theo s xut hin mt t bo ch cha nhim sc th ti t hp. Tu theo cch chuyn vn ADN m ta phn bit ba kiu chuyn tnh trng vi khun: bin np, ti np v tip hp. Ti t hp di truyn: C hai c ch qua ADN l xm nhp vo t bo vi khun c th ti t hp vo nhim sc th vi khun hoc plasmid: ti t hp ph bin hoc tng ng v ti t hp c hiu v tr hoc c hiu th t. 4.1.1. Ti t hp ph bin Ti t hp ph bin l qu trnh m trong ADN l mi xm nhp vo trong t bo c lin kt vi ADN ca t bo ch qua vic ghp i on tng ng, b v v trao i cho 2 on ADN c trnh t ging nhau. t nht c 6 enzym tham gia vo qu trnh ny, trong ng ch l protein lin kt si n SSB protein, v RecA protein (hnh 5.6).
ADN t bo nhn
ADN t bo nhn ti t hp
Hnh 5.6. C ch ti t hp ph bin
4.1.2. Ti t hp c hiu v tr Ti t hp c hiu v tr hoc c hiu th t xy ra ti v tr c bit, ch cn mt on ADN
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tng ng rt nh nhn bit gi l trnh t nhn bit, qu trnh ny cn cc enzym c hiu cho cc phn t ADN ti t hp. Nu ch mt phn t ADN mang th t nhn bit, th gi l ti t hp c hiu mt v tr. C hai phn t ADN u mang trnh t nhn bit, th gi l ti t hp c hiu hai v tr. S hp nht ca phage v ca plasmid F vo genom ca E. coli l v d v kiu ti t hp ny (hnh 5.7).
Hnh 5.7. Ti t hp c hiu v tr nh phage vi: AATP v tr gn ca phage Attachment site of phage ATTB v tr gn ca vi khun Attachment site of bacterium
Khi chuyn sang trng thi prophage, phage lambda () gn vo nhim sc th ch v tr gia galactoseoperon v vng biotin. Vic gn m u bng s sp hng ca hai th t base tng ng gm 15 cp base dng lm vng nhn bit i vi enzym integrase do phage tng hp. Enzym ct ADN si kp ca phage v ca t bo ch vng ny (ATTP v ATTB). Cc u t do xut hin v s c lin kt cho nh ligase. S lp vo ca phage l thun nghch. Mt enzym khc cng do phage c m, excisionase xc tc vic tch genom ca phage ra. C hai qu trnh lp vo v tch ra ca phage ny din ra c s tham gia ng thi ca mt protein to thnh bi t bo ch, nhn t hp nht ca t bo ch, IHF (integration host factor). 4.1.3. Cc nhn t di truyn ng Cc nhn t di truyn chuyn dch ch (IS, Transposon, phage Mu) tng cng kh nng ti t hp di truyn vi khun. 4.1.3.1. Nhn t chn vo IS (insertion sequence) IS l cc nhn t di truyn, c th lp vo cc v tr rt khc nhau trn genom ca vi khun. c tnh ny to cho nhn t IS kh nng di ng rng ri, cn c gi l chuyn ch. Cc nhn t IS c tm thy u tin cc bin chng ngu nhin E. coli, do chn vo ADN ca t bo ch m lm mt tnh lin tc ca cc gen. Chng gm khong 800 1.400 cp base nucleotid v khng mang mt phenotyp no khc ngoi chc nng chuyn ch. Chc nng ny c m bo bi enzym transporase. 4.1.3.2. Transposon (Tn) Cng thuc cc nhn t di truyn ng, gy t bin v cng c gi l gen nhy. Khc vi nhn t IS, cc Tn c m cho mt s tnh trng d nhn thy v kiu hnh nh tnh khng khng sinh (penicilin, tetracyclin, kanamycin,...), tnh khng kim loi nng (v d Ag). S nhy hoc s chuyn ch ca Tn l kt qu ca s sao chp Tn m khng lm mt Tn v tr chn vo ban u trong ADN. 4.1.3.3. Bacteriophage Mu Bacteriophage Mu c chung c tnh hp nht khng bnh thng ca IS v Tn. Khi hp nht vo gen ca t bo ch, phage s gy t bin (Mu l vit tt mutatortc nhn t bin).
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Bacteriophage Mu tc dng nh Tn khng l. o ngc gen l c tnh ng ch ca Bacteriophage Mu. S o ngc u on G ca Bacteriophage Mu. on G m ho tng hp cc protein si ui. Nu on ny nm theo hng G+, cc protein ui S v U c to thnh v phage nhim vo E. coli K12. Nu on o ngc theo hng G, mt promotor khc s hot ng v si ADN i din s c s dng, si ny phin m cc gen khc m sn phm ca chng l mt protein ui S v U khc lm thay i ph vt ch ca phage. Nh vy, phage c th hp th vo cc t bo E. coli C v cc vi khun ng rut khc. S o ngc cng lin quan n Salmonella, v y lin quan n s thay i tng hp tin mao ca vi khun.
4.2. Bin np (transformation)

S chuyn gen ADN c gii phng t mt vi khun cho hoc chit c t vi khun ny sang vi khun nhn khc c gi l bin np. Cc t bo trng thi c th bin np c bi mt ADN trong mi trng l cc t bo kh np (competent). Cc chi Acinetobacter, Azotobacter, Bacillus, Haemophyllus, Mycobacterium, Neisseria, Pseudomonas, Streptococcus v Synechococcus c c tnh ny. Trng thi kh np do cc gen nhim sc th m ho v c kch thch bi mt s iu kin mi trng. Cc vi khun nh th l cc vi khun c kh nng kh np t nhin. Nhiu vi khun phi qua x l nhn to, v d: vi cc cation ho tr 2 nng cao mi tr thnh kh np, gi l cc h thng bin np nhn to. i vi cc D. pneumoniae, khi khong 12 protein phn t nh c tng hp v c bi tit vo mi trng, v khi nng ca c t bo v yu t kh np t n nng nht nh th tnh kh np c pht trin. Mng ngoi ca thnh t bo kh np cha mt s cu trc dng ti l nhng np gp mng ngoi, gc ca mi ti c nhng capilaire nh. Bn trong mi ti c protein lin kt c hiu vi mt trnh t ADN gm 11 cp base bt gp 600 v tr trn genom. Ngay sau khi ADN tng ng c b sung vo qun th vi khun kh np, cc ti hng ra ngoi bin mt v xut hin cc ti hng vo trong. C th s lin kt ADN lm ti b mt gp khc, to thnh cu trc c gi l transformasom. ADN bn trong transformasom khi i qua thnh t bo ch c chuyn thnh dng si n ngay trc khi ti t hp vi genom t bo nhn. Cn t bo H. influenzae c th hp th cc plasmid nguyn vn nu plasmid cha trnh t 11 cp base ph hp. B. subtilis khi bin np vi ADN nh ca D. pneumoniae v plasmid bao gi cng phn ct v chuyn chng thnh si n khi xm nhp t bo. cc vi khun kh np t nhin, tnh trng c bin np l tnh khng c t v tnh nguyn dng i vi acid amin. Trong trng thi sinh l thch hp, t bo kh np c b mt t bo thay i, thnh t bo xp v c hot tnh ca cc enzym ngoi bo cao, ng thi yu t kh np c to ra v tit vo mi trng. Nng ca ADN thch hp cho bin np ch cn thp: vi 0,1g ADN/ml huyn dch t bo l c th bin np 5% qun th t bo nhn. i vi cc vi khun trong iu kin bnh thng khng c kh nng bin np, tnh kh np c th cm ng c nh x l t bo vi dung dch CaCl2 hoc nhit lnh. i vi cc vi khun G+ chi Bacillus v cc x khun chi Streptomyces, vic s dng cc th nguyn sinh cht thiu vch (t bo trn) bin np c ngha thc t cao. Vic hp th ADN plasmid nh cm ng vi polyethylenglycol qua th nguyn sinh cht thiu vch, hay chuyn ADN nhim sc th qua dung hp trc tip cc th nguyn sinh cht c th thc hin c. Cc th nguyn sinh cht dung hp kt hp genom ca c hai t bo b m c th ti sinh li thnh cc t bo nguyn vn trong cc iu kin thch hp. Cc th ti t hp c to ra t s dung hp nh trn mang tnh trng ca c hai b m nh qu trnh ti t hp. Phng php dung hp t bo trn (lai ghp nguyn sinh cht) nh xung in cng c s dng c hiu qu. Phng php ny to cho cc mng sinh hc tr thnh d thm v d dung hp nh xung kch thch ca in trng. Cc ADN plasmid ca vi khun G+ v G cng c th bin np thnh cng nh xung in. Th nghim Grifith l th nghim bin np kinh in (Xem gio trnh Sinh hc i cng).
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4.3. Ti np (transduction)
Ti np l s chuyn gen ADN t t bo cho sang t bo nhn nh phage. Thng ch mt on ngn ADN ca t bo cho c chuyn. C ti np ph bin v ti np c hiu v trong c hai trng hp phage ti np thng b khuyt tt, nh mt kh nng dung gii t bo ch. Ti np gp cc loi Bacillus, Escherichia, Pseudomonas, Rhizobium, Salmonella, Shigella, Staphyllococcus v Vibrio. Tuy nhin, khng phi tt c cc phage u c th ti np v tt c vi khun u ti np c. 4.3.1. Ti np khng c hiu (ti np ph bin) Ti np khng c hiu l kiu ti np m trong mt on bt k ca ADN t bo ch c lp thm vo hoc thay th bng genom ca phage. Ti np khng c hiu c Lederberg v Zinder pht hin nm 1951 S. typhimurium. Chng cho S. typhimurium B+ c nhim pha n ho P22. Sau khi t bo ch b dung gii, cc pha c tch ring v c vi chng nhn S. typhimurium B khc vi S. typhimurium B+ t nht mt c tnh di truyn. Khi cy sng lc trn mi trng chn lc nhn thy trong s cc S. typhimurium B c mt s bin th c tnh trng ca S. typhimurium B+. Nh vy khi phage P22 nhn ln trong S. typhimurium B+, ADN ca t bo ch b phn ct thnh cc phn on, v mt vi on no c th c bao bc bi v capsid ca phage. Tt nhin, trong s cc phage mi c to ra vn c cc phage bnh thng do m dn n s dung gii t bo. n giai on sau, trong s cc t bo S. typhimurium B nhn phage, mt s nhn c phage ti np khuyt tt c ADN ti t hp vi h gen ca th nhn. Cc on tng ng trao i c th a n vic thay th mt gen khuyt tt trong th nhn bng mt gen nguyn vn ca th cho. V ch c mt on nh ADN ca th cho c ti np, nn xc sut ti t hp ca mt tnh trng nht nh l khong 108 106. i vi phage P22 ca Salmonella v phage P1 ca E. coli, ch mt tnh trng hoc cc gen nm rt gn nhau l c th c ti np. on ADN c ti np vi ADN ca phage ch bng 1 2% ADN ca vi khun. c bit, phage PBS1 ca B. subtilis l ngoi l khi c th ti np c 8% genom ca t bo ch. 4.3.2. Ti np c bit Trong trng hp ny, ch mt s on ADN xc nh c ti np m thi. Chng hn, phage thng ch ti np gen gal v gen bio. Nu phage ti np nhim vo mt t bo nhn b khuyt tt, chng hn gen gal (gal), ti t hp c th xy ra qua trao i gen gal bng gen ti np gal+. Cc th ti t hp hoc cc th ti np to thnh s l gal+. Vic ti np gen bi phage 80 cng ging th: phage chn vo ngay cnh gen trp, do ti np c gen trp. Trong ti np c hiu th s chn c hiu ca phage vo genom ch l tin quan trng cho vic ti np ADN t hiu qu. Trong mt s trng hp, on ADN ti np khng ti t hp m nm ngoi nhim sc th ca th nhn: t bo l d hp v tnh trng c chuyn ti ADN ti np c phin m nhng khng c sao chp. V vy, khi phn bo, on ADN ca th cho ch c phn vo mt t bo con. Nhng t bo nh th c gi l th ti np khuyt. Nu th nhn l khuyt dng v tnh trng v ADN ca th cho b sung cho tnh trng y, th ch nhng t bo di truyn tip on ADN ni trn mi c th sinh trng v to thnh khun lc rt nh trn mi trng thch. Hnh 5.8 minh ho qu trnh ti np.
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4.3. Tip hp (conjugation)

Tip hp l s vn chuyn ADN qua thit lp cu tip hp trc tip gia hai t bo vi khun, s chuyn dch c nh hng t t bo cho (c) sang t bo nhn (ci) (hnh 5.9).
Hnh 5.9. Tip hp vi khun
T bo cho cha mt yu t ADN c th di chuyn c gi l plasmid gii tnh F (fertility plasmid). Nhng t bo thiu plasmid F (F) ch c th l th nhn. Khi tip hp, plasmid F c
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chuyn vi xc sut 100% nhng khng tnh trng no ca nhim sc th c hp chuyn. Plasmid F c th hp nht vo nhim sc th, khi tip hp, ADN ca nhim sc th s c chuyn t vi khun cho sang vi khun nhn vi tn sut cao hn hng ngn ln so vi dng chng F+. Cc t bo ny c gi l cc t bo Hfr (High Frequency of Recombinants = Tn sut cao ca cc th ti t hp). Plasmid F lp thun nghch vo nhim sc th v tr xc nh, v khi tch ra nu khng chnh xc s ko theo mt on ADN nhim sc th t bo, to thnh plasmid F (F = F + on ADN). T bo cha F gi l t bo F.
Plasmid F l phn t ADN si kp, vng kn, kch thc khong 105 base, c kh nng t sao chp c lp vi nhim sc th. Plasmid F cha cc gen cn cho s tip hp, cc gen xc nh tim mao gii tnh F. N c mt c tnh chung ging vi cc plasmid khc nh cha mt s gen cho php sao chp trong t bo, th hin tnh khng tng thch do vng Inc quy nh: nu c mt plasmid trong t bo th s sao chp ca cc plasmid thn thuc s b km hm. Plasmid F thuc nhm khng tng hp gi l Inc F1 (hnh 5.10).
Cc plasmid thng thng t gp cc procaryota. Cho ti nay mi ch pht hin c Streptomyces, Streptococcus v Nocardia opaca. S tn ti ca chng trong t bo trnh c s phn gii ca exonuclease cn cha r.
4.4. Vai tr ca cc plasmid trong sinh hc

Cc tnh trng c m ho bi plasmid thng cung cp cho cc t bo ch cc u th sinh trng, v nh m cc t bo ny thu c u th chn lc. 4.4.1. Plasmid khng Nh trnh by, plasmid R (plasmid khng thuc) l nguyn nhn ca khng thuc ngoi nhim sc th, gip cho vi khun tip nhn khng vi cc khng sinh v ho tr liu. Mt s
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plasmid khc cung cp tnh khng vi kim loi nng, nh bc, nicken, coban, cadimi, ng, km, crm, arsen, antimon, telua hoc thu ngn. Cc plasmid R c th c chuyn ti nh tip hp hoc bin np. Cng vi s chuyn ti ca plasmid R, mt s gen nhim sc th cng c chuyn, nh vy cc gen ny c huy ng bi plasmid R. 4.4.2. Plasmid m ho bacteriocin Nhiu vi khun to thnh cc protein c kh nng git cht hoc km hm sinh trng ca cc loi thn thuc. Cc protein c tc dng c hiu ny c gi l cc bacteriocin v do plasmid c m. Bacteriocin c phn lp t E. coli (colixin), P. aeruginosa (pyoxin), B. megaterium (megaxin),... 4.4.3. Plasmid m ho yu t gy bnh Cc VSV c kh nng nhim hi v chng c cc yu t xm nhim, yu t gy bnh v cc yu t c tnh. Mt s trong s cc yu t ny do plasmid c m, cng v vy m cc yu t y c ph bin nhanh chng. l cc yu t xm thc gip VSV pht trin mnh m trn b mt cc mng nhy (invasin), c plasmid c m gip cho Shigella flexneri xm nhp tch cc vo trong cc t bo biu m rut. Enterotoxin (c t rut), c tc dng c trong ng rut v gy a chy, thng cng c c m bi plasmid. Hemolyzin c tc dng dung gii hng cu. i vi cc vi khun ng rut gy bnh, kh nng s dng ion st l tin quan trng cho s xm nhp ca chng vo cc t bo v m. V cc siderophor lin kt st nh aerobactin cng c c m bi plasmid. 4.4.4. Plasmid m ho trao i cht phc tp C mt s plasmid mang gen chu trch nhim cc phn ng trao i cht phc tp: nh c nh nit to thnh nt sn, tng hp cc enzym ca qu trnh phn ng nitrat ho,.... Cc plasmid trao i cht ny thng l cc plasmid khng l, kch thc khong 300 1.200kb, c gi l cc mega plasmid. c bit, mt s h thng gii hn v ci bin bo v vi khun khi s xm nhp ca ADN l mt s vi khun c ngun gc t plasmid, tuy nhin, trong s vi khun khc gn gi v chng loi pht sinh li nm trn ADN. S nh v khc nhau ca gen cho thy h gen gia nhim sc th v plasmid c trao i v plasmid ng vai tr quan trng trong tin ho ca h gen t bo nhn nguyn thu. Hnh 5.11 gii thiu bn nhim sc th ca E. coli.
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Hnh 5.11. Bn nhim sc th ca E. coli
4.4.5. Cc th h plasmid Cc plasmid th h th nht: y l cc plasmid tm thy trong t nhin (ColE1, pSC101,...), gp phn trong to dng u tin cc gen Eucaryota. Nhng cc plasmid ny c t cc c tnh cn thit nn cc nh nghin cu to ra cc plasmid th h hai. Cc plasmid th h hai: y l cc plasmid c to ra bng cch tp trung cc c tnh qu ca nhiu plasmid t nhin vo mt plasmid. H plasmid pBR c hnh thnh theo cch ny, m i din rt ni ting l plasmid pBR322. pBR322 c kch thc 4.363bp, mang 2 gen khng khng sinh (khng ampicilin v tetracyclin) v 20 v tr nhn bit ca cc restrictase (RE), m 11 trong s nm trong cc gen khng khng sinh, nn vic gn mt trnh t ADN l vo mt trong cc v tr s lm mt tnh khng thuc tng ng. Cc plasmid th h ba: y l cc plasmid mnh nht hin nay vi 2 c tnh c bn l: (1) kch thc nh nn sao chp rt nhanh, to c nhiu bn sao trong vi khun; (2) c mang mt polylinker (polycloning site), l mt on polynucleotid tng hp tng ng vi mt dy cc v tr nhn bit duy nht ca cc RE. C 3 phn nhm chnh: + Cc plasmid nhm pUC: khong 2.600bp, c gen ampr v mt phn gen lacZ, xen vo gia lacZ l polylinker. + Nhm cc plasmid pSP v Gemini: khong 3.000bp, c gen ampr v polylinker, khng c lacZ, nhng cc plasmid nhm ny (pSP64, 56, Gemini) c promotor c trng cho ARN polymerase (SP6, T7) cho php phin m ADN gn trong vector thnh rt nhiu on ARN,
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+ Cc plasmid Bluescript: Kt hp c u im ca cc nhm trn v c tim nng nht hin nay.
5. GII HN V CI BIN V C S K THUT GEN

Nhn chung phage nhim sinh pht trin trong chng E. coli ny li khng th nhim sinh trong nhng chng khc. S hn ch ny chnh l do enzym ca vi khun nhn ra v tr c bit trn ADN l ca phage v ct loi b chng, khi ADN ca t bo c bo v nh s ci bin bi enzym v do khng b enzym gii hn (restrictaseRE) nhn ra. H thng gii hn v ci bin l kh ph bin VSV, mt mt c s dng nh du ADN ca t bo, mt khc li phn hu ADN l mi xm nhp. H thng enzym gii hn (restrictase) gm c hai loi hot tnh enzym: endonuclease v methyltransferase. C hai th hin hot tnh mt on xc nh th t phn bit. Trong trng hp ca ADN t bo, th t ny c ci bin ho hc nh methyltransferase qua phn ng methyl ho adenyl v tr N6 v khng b ct bi endonuclease qua s thu phn lin kt phospho dieste. Mt trnh t nucleotid xc nh l c cht cho mt hoc hai enzym hot tnh m thi. Methyltransferase ci bin ADN ca t bo khi sao chp, sn phm mt si kp ADN bi si n ADN c methyl ho. Sau , hot tnh endonuclease c kch thch khi trong t bo xut hin ADN khng b ci bin (ADN l), l c cht cho enzym ny.
5.1. Cc endonuclease gii hn

Cc endonuclease c dch m bi genom vi khun, phage v plasmid. C ba loi endonuclease v u ct ADN si kp. Enzym gii hn Loi I c cng mt protein mang c hai hot tnh (endonuclease v methyltransferase) c trnh t nhn bit c bit, ct ADN ngu nhin. Enzym gii hn Loi II c endonuclease v methyltransferase c mang bi hai protein khc nhau, c trnh t nhn bit c hiu, v tr ct cng c hiu v nm trong vng nhn bit, th hin s i xng quay kp (hai si c cng trnh t khi c theo cng mt chiu phn cc). Kt qu l xut hin nhng on ADN vi trnh t xc nh. Endonuclease loi II c s dng hu hiu trong k thut to dng nhn bn gen. Enzym gii hn loi III c cng mt protein mang c hai hot tnh (endonuclease v methyltransferase), c v tr nhn bit c hiu v v tr ct cng c hiu. Hin nay c khong hng trm enzym gii hn c thng mi trn th trng (bng 5.1).
Mi tn ch v tr ct ca enzym, ng ------ l trc quay th t nhn bit, v ngi sao * ch methyl ho. Nhiu enzym endonuclease ct 2 si ADN trn trnh t c hiu lch i s to ra cc on ADN
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gn u dnh (c kt), cn s enzym khc nu ct trn cng mt v tr s to ra cc u bng (t). Trn cc on ADN vi cc u dnh, cc si n b sung c th gn vi nhau nh lin kt hydro, sau nh ligase kt hp vi s dng ATP cc lin kt ng phosphat s c hn li.
5.2. Thit k cc plasmid (vector) ti t hp

Nu hai loi ADN c x l vi cng mt enzym gii hn (restrictase) th mi ADN s c u dnh tng hp nhau v 2 on ADN y s gn li c vi nhau to thnh ADN duy nht. Plasmid m c cha 2 loi ADN c ngun gc khc nhau c gi l plasmid ti t hp. C th gn gen (bn sao cADN) ca sinh vt nhn tht bc cao (v d: vn, ngi) vo mt plasmid, nhng cn thc hin theo cc bc sau: t m vt ch ta chit ly premARN t gen mc tiu, ri loi b cc intron v ghp ni cc exon ta c mARN trng thnh. T mARN trng thnh nh enzym phin m ngc tng hp cADN (k thut RTPCR), v cADN c ghp vi plasmid to thnh plasmid lai (hnh 5.12).
5.3. a plasmid ti t hp vo t bo vi khun nhn

Phng php a plasmid ti t hp ny vo t bo thng s dng l phng php bin np. Tuy nhin, a s vi khun khng d dng tip nhn plasmid ti t hp. tng cng tn sut bin np ngi ta x l t bo nhn vi dung dch CaCl2 nhit thp (0 4OC vi E. coli), b sung plasmid ti t hp ri un nng t bo nhn (ti 42OC vi E. coli) gy shock nhit chung. Nh vy, tnh thm ca t bo VSV thay i, cho php plasmid xm nhp t bo. Plasmid cng c th c t bo hp th sau khi b x l vi shock in (xung in), shock in to thnh cc mao dn, qua plasmid c
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th d dng xm nhp t bo. Ngi ta s dng cc gen khng khng sinh nh khng tetracyclin, ampicilin lm du hiu nhn bit t bo dung np plasmid. Nu ban u cc vi khun nhn l mn cm vi cc khng sinh trn th sau khi bin np trong qun th VSV, nhng t bo no khng khng sinh chnh l nhng t bo tip nhn plasmid bin np. Khi hn dch cc vi khun sau bin np c cy sng lc trn mi trng thch c cha khng sinh ch th, th ch nhng vi khun c cha plasmid ti t hp mi pht trin to thnh khun lc c. Khi t bo vi khun nhn plasmid ti t hp, t bo sinh sn to thnh mt qun th ln cc t bo ging nhau gi l mt dng. V plasmid l tc nhn bin truyn nh m gen mi c a vo t bo vi khun nn plasmid cn c gi l vect (vector) to dng. Vi mt s plasmid, vic cho vo mi trng nng thp ca chloramphenicol s dn n vic sao chp plasmid mt iu khin khin cho hng trm bn sao plasmid (v tt nhin gen ngoi lai trong ) c th c tng hp bi mi mt t bo vi khun. S sao chp bt thng nh vy ca plasmid gi l s khuch i.
6. K THUT PCR (POLYMERASE CHAIN REACTION) V XC NH TRNH T ADN
6.1. Phn ng chui trng hp (PCR)

Phn ng chui trng hp PCR l phng php to dng (khuch i) ADN in vitro, c p dng rng ri trong k thut di truyn ngy nay. Cng b trong nm 1985 nh pht minh ca Karl Mullis v cng s, k thut PCR thc s tr thnh cng c c lc ca sinh hc phn t v k thut di truyn. 6.1.1. Nguyn tc
Trong iu kin in vitro, vi cc on mi chuyn bit bt cp c vi hai u mt on ADN, enzym Taq ADN polymerase, ion Mg2+ v cc nucleotid tng ng, trong 3 chu k nhit thch hp, on ADN nu s c tng hp khuch i ln mt cch mnh m. Tt c cc ADN polymerase khi hot ng tng hp mt mch ADN mi t mch khun u cn s hin din ca nhng mi chuyn bit. Mi (on ADN mi probe) y l nhng on ADN ngn c kh nng sng i b sung vi mt u ca mch khun v ADN polymerase s ni di mi hnh thnh mch ADN mi, nm gia hai mi. khuch i mt trnh t ADN mc tiu, cn c thng tin ti thiu v trnh t to cc mi b sung chuyn bit. Cc mi ny bao gm mt mi xui (sens primer) v mt mi ngc (antisens primer). Xui v ngc c hiu theo ngha xui v ngc so vi chiu phin m ca gen.
6.1.2. Thc nghim Phn ng PCR bao gm chui phn ng nhiu chu k ni tip nhau. Mi chu k gm 3 bc sau (hnh 5.13): Bc 1. Trong dung dch phn ng c cc thnh phn cn thit cho sao chp, phn t (on) ADN c bin tnh 94 95OC (nhit ny cao hn nhit nng chy Tm ca ADN) trong 30 giy 1 pht. Si kp ADN s tch thnh 2 si n. y l giai on bin tnh (denaturation). Bc 2. Nhit lc ny c h xung (thp hn Tm ca cc mi) cho php cc ADN mi bt cp c vi cc ADN khun si n. Chu k nhit ny giao ng trong khong t 38 70OC, tu thuc vo Tm ca cc mi c s dng, v ko di t 30 giy 1 pht. y l giai on lai.
Bc 3. Nhit c tng ln 72OC, v c gi y t 30 giy vi pht gip cho ADN polymerase chu nhit (Taq ADN polymerase) tng hp cc ADN si p mi t cc ADN si n c bt cp vi on mi t chu k trc. y l giai on tng hp hay ko di.
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3' 5' Lp 3' 5' li B 3' n ln 5' C 3' 5' 72OC 3870OC A 9495OC
5' 3'
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Hnh 5.13. Cc bc chu k ca k thut PCR
A. Bin tnh: tch ri 2 mch ca phn t (on) ADN; B. Lai: cp mi chuyn bit cho mt trnh t ADN c cho bt cp vi khun; C. Ko di: ADN polymerase tng hp mch ADN mi t mi bt cp. Chu k ny c lp li n ln.
Mt chu k gm 3 bc trn s c lp i lp li nhiu ln v mi ln s lm tng gp i lng mu ADN ca ln trc. y l s khuch i theo cp s nhn: sau 30 chu k, s khuch i s tng ln 106 ln so vi lng khun mu ADN ban u. ADN mu tht tinh khit l tt nht cho PCR, tuy nhin, ADN thu nhn c trc tip t dch chit t bo vn c th s dng c. Lng ADN mu c th trong khong 100ng n 1g. ADN polymerase s dng y l enzym chu nhit chu nhit Taq ADN polymerase c chit xut t vi khun sui nc nng Thermus aquaticus. Ngy nay, bn cnh Taq ADN polymerase cn c enzym Tth ADN polymerase cng c ng dng (c chit t Thermus thermophylus), hot ng nh enzym phin m ngc khi c mt ca ARN lm khun v ion Mn2+; tuy nhin khi c mt ADN khun v ion Mg2+ li hot ng nh ADN polymerase. Vy khi khuch i ARN, s dng Tth ADN polymerase cn b sung Mn2+, kt qu s thu c cADN t ARN khun ban u. y l nguyn l c bn ca phng php RTPCR rt hu hiu khuch i cc mARN, vrARN,... Mi phi c chn khng c sng i b sung gia mi xui v mi ngc, Tm khng cch nhau qu xa, c cn bng cp base, c tnh c trng cho ADN cn khuch i. Nng 4 loi nucleotid thng l 20 200M cho mi loi. Nng Mg2+ cng rt quan trng, tuy nhin phi xc nh c th. Thc t khng vt qu 40 chu k cho mt phn ng PCR. Nu s lng bn mu ban u l 5, th cn 20 30 chu k. Nu s lng mu l 102 103, cn 35 40 chu k. 10
6.2. K thut xc nh trnh t ADN

Hai phng php xc nh trnh t chnh l phng php ho hc ca Maxam v Gilbert (1977) v phng php enzym hc qua vic s dng cc dideoxynucleotid ca Sanger v cng s (1977). V tnh tin ch v n gin bt, y ch trnh by phng php ca Sanger v cng s. Phng php Sanger da vo s tng hp nh enzym ADN polymerase mch b sung cho trnh t ADN mch n cn xc nh. c trng ca phng php l ngoi 4 loi nucleotid thng thng
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cn cn s dng thm 4 loi dideoxynucleotid khng c kh nng hnh thnh cc lin kt phosphodiester v v th lm ngng qu trnh tng hp. Trnh t ADN cn xc nh phi c to dng trong mt vector mch n (phage M13). ADN polymerase s dng y c th l on Klenow ca ADN polymerase hay Taq ADN polymerase, hay Sequenase. S tng hp mch mi bt u t 1 mi sng i vi trnh t chuyn bit trn phage M13, vi s hin din ca 4 loi nucleotid, trong 1 loi c nh du ng v phng x (35S). Phn ng tng hp c tin hnh trong 4 phn on ring. Ngi ta ln lt cho vo mi phn on mt trong 4 loi dideoxynucleotid (ddNTP) vi hm lng rt nh (~1% so vi dNTP). Do nng thp m thnh thong mi c mt dideoxynucleotid c s dng vo phn ng tng hp mt olygonucleotid; v lp tc, s tng hp olygonucleotid ngng li. Tnh xc sut th trong mi phn on s c mt tt c cc nucleotid hin din trn trnh t ADN i tng. V d, nu trnh t ADN cn xc nh l AATCGATAGGCTTGCATG th trong phn on c mt dideoxynucleotid ddC s c s tng hp cc olygonucleotid sau: AATC AATCGATAGGC AATCGATAGGCTTGC AATCGATAGGCTTGCATG Sau , 4 phn on phn ng tng hp s c em phn tch trn gel polyacrylamid v kt qu c c trn bn phng x t ghi (hnh 5.14).
Hnh 5.14. Phng php Sanger s dng cc dideoxynucleotid
Trnh t cn xc nh c to dng trong phage M13. V d cho thy s hnh thnh 5 vch trong bn in di c cho thm ddATP; mi vch tng ng vi mt on ADN kt np mt ddA thay v dA v do khng tip tc ko di thm. Bn in di c c theo chiu mi tn. Trnh t c bn tri chnh l trnh t b sung cho trnh t c c trn bn in di.
Tuy nhin, c mt s ci tin n gin ho phng php trn. Ngi ta to dng ADN cn
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xc nh trnh t vi vector l plasmid th h 3 ngay t u. hai bn ca on ADN c to dng, cc plasmid ny c mang hai trnh t chuyn bit, mi trnh t nm trn mt mch. Khi cn xc nh trnh t ca mch no ngi ta bin tnh tch ri hai mch (nh NaOH) ri s dng mi bt cp vi trnh t chuyn bit nm trn mch . Phn ng tng hp xy ra theo nguyn tc nu trn. Khi xc nh trnh t bng my t ng, ngi ta khng nh du bng cc ng v phng x m bng ho cht cc flourchrom. Mi loi dideoxynucleotid c nh du bng mt flourchrom c mu khc nhau. Nh vy, tt c cc olygonucleotid cng kt thc ti mt loi dideoxynucleotid s c cng mt mu. Sau khi in di trn gel polyacrylamid, kt qu s c c qua h thng gn vi tnh iu khin. PCR cho php xc nh trnh t mt ADN c khuch i bng PCR khng qua to dng. Phng php ch s dng c khi trnh t on ADN c bit trc, c ng dng xc nh nhanh t bin im. Trc ht, on ADN cn xc nh trnh t c khuch i bng phng php PCR. Sau , hai mch ca ADN c tch ri ra. Mi mch c bt cp vi mt mi (c th l mi dng cho phn ng PCR hay mt mi nm bn trong on ADN). Phn ng tng hp xy ra tng t nh enzym s dng dideoxynucleotid.
7. CC P DNG CA K THUT GEN (GENETIC ENGINEERING)

K thut gen k ngh di truyn hin ti v tng lai c th gp phn quan trng vo chm sc sc kho, cung cp dc phm, thc phm, cung cp dch v, ci to mi trng. Cng ngh dc phm s dng k thut bin i gen sn xut c cc hot cht, dc cht dng trong iu tr nh: insulin ngi, yu t sinh trng ngi (hGH), plasminogen ca m, urokinase (dng x l cc mu ng), interferon, somatostatin (hormon v no),... Cc cht hot ng sinh hc c sn xut nh k thut gen ngy cng nhiu v ng dng ngy cng a dng. K thut mi sn xut vaccin c pht trin t k ngh di truyn.
7.1. Vaccin phng bnh l mm long mng, vaccin ti t hp v vaccin chng vim gan B
Gen protein v c hiu ca virus gy bnh l mm long mng ng vt c a vo plasmid v plasmid ti t hp c bin np vo E. coli. Khi nui cy E. coli bin i gen s to ra protein c th trn, v sau khi c chit xut ri tim vo tru, b s kch thch sn xut khng th trong ng vt chng li bnh l mm long mng. Phng php sn xut vaccin ny cho php trnh c nguy him v kh khn gp phi khi sn xut vaccin bnh l mm long mng s dng phng php thng thng. Trong trng hp vi khun Vibrio cholera, ngi ta loi b mt phn gen m ho enterotoxin ca vi khun trc khi dng chng sn xut vaccin. Kt qu l protein sinh ra c hot tnh gim ng k m vn gi c tnh khng nguyn cao. Kt qu tng t cng nhn c i vi S. typhimurium vi t bin trn gen aro. Mu ca nhng ngi mc bnh virus vim gan B (HBV) kinh nin c cha cc ht protein nh v hi ca virus. y l protein HBsAg, c th chit xut c, ri x l dit virus v tinh ch. Khi tim HBsAg vo trong ngi s kch thch min dch chng li cc virus gy bnh. Mt khc HBsAg c th sn xut c nh S. cerevisieae bin i gen (tip nhn gen m ho cu trc ca HBsAg), v HBsAg ni bo c chit xut sau khi t bo nm men b ph v. Vi k thut ny, protein HBsAg bin i gen c sn xut vi khi lng ln v s dng lm vaccin phng bnh vim gan B. y l vaccin bin i gen u tin c sn xut phng bnh cho ngi.
7.2. Vaccin chng AIDS

Gen m ho gp160 ca HIV c gn vo ADN ca Bacilovirus tn cng cn trng. Khi cc t bo m cn trng b nhim Bacilovirus ti t hp c nui cy, gen gp160 c biu hin v protein gp160 c to ra. Th nghim v to ra p ng min dch ch ng ngi i vi protein gp 160 ang c nghin cu. Tuy nhin cc t bin khng vi tn sut xut hin cao l tr ngi
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ln trong nghin cu pht trin vaccin chng HIV/AIDS. Vaccin ADN HIV1 v ang c th nghim phng chng HIV/AIDS. Vaccin ADN HIV1 l vector liu php gen ci bin t b gen ca HIV1. Mt s loi vaccin ADN th nghim phng chng HIV/AIDS c to thnh t cc plasmid ti t hp gm pTXGE v cc gen c hiu ca h thng min dch ngi nh il2, ifn v gen gmcsf. Plasmid th h 3 pTXGE c to thnh t plasmid pTX (4,55kb) gn thm b gen HIV1 ci bin ( xo b mt s vng gen ca HIV nh gen pol, v cc gen vpu, vpr, tat b xo mt vi vng nh). Vaccin ADN HIV1 th nghim trn ng vt thc nghim cho kt qu tt, c th l mt trong nhng gii php chng HIV/AIDS trong tng lai.
7.3. Sn xut insulin

Vi s pht trin mnh m ca cng ngh gen, ngi ta c th nh VSV sn xut c cc hot cht m khi thu khng mt thng tin no c cha trong VSV sn phm hon ton xa l vi VSV. Nguyn l: Gen chuyn trch v cu trc insulin thu c t cc ngun khc nhau (tng hp ho hc, hay phn lp t cc ngun khc nhau ri c khuch i) c a vo t bo VSV. Khi cc VSV bin i gen ny c nui cy cho pht trin s biu hin gen mi ny v to ra hot cht insulin m ta mong mun. Insulin bao gm 2 mch polypeptid: mch di 30 (B) v mch ngn 21 (A) n v acid amin. Gen m ho polypeptid ny cha hai chui ADN: chui di cha 18 phn on nucleotid, v chui ngn cha 11 phn on. Hai chui nucleotid ny c tng hp thnh hai on gen ring bit v c ghp ring r vo plasmid pBR322 gn phn cui ca gen galactosidase. Plasmid c bin np vo E. coli v c nhn bn ln trong E. coli. Sau khi c phin m sang mARN, hai mch A v B ca insulin c to thnh v c bi tit ra cng vi galactosidase. Sau khi tch tng polypeptid khi enzym bng BrCN, hai mch A v B c gn li vi nhau to ra insulin hot ng (k thut 2 block, hnh 5.15).
Gly Ile Val Glu S Thr Ser S Cys Gly Ser His Leu Val Glu Ala Leu Tyr Leu Val S Ile Cys Ser Leu Tyr Gln Leu Glu Asn Tyr Cys S S Cys Gly Chui B Glu Arg Thr
Insulin ngi
Chui A
Gln Cys Cys S His Gln Asn Val Phe Leu
Asn
Lys
Pro
Thr
Tyr
Phe
Phe
Gly
Phng php sn xut insulin mt bc cng bng k thut ti t hp ADN, cc nh khoa hc gn gen m ho tng hp proinsulin vo t bo E. coli, ri nui cy E. coli bin i gen v chit ly proinsulin. Thu phn tin insulin bng protease ct mch peptid C (cha 34 acid amin) cho ta insulin hot ng. Phng php ny cng c th p dng c i vi nm men S. cerevisiae (hnh 5.16).
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Hnh 5.15. Sn xut insulin vi k thut 2 block
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7.4. K thut di truyn vi sinh vt vi nng nghip

Trong nng nghip ngy nay c th s dng cc chng Pseudomonas syringae v Pseudomonas fluorescens bin i gen bo v cy qu chng li sng gi. Nh k thut ADN ti t hp, mt phn gen m ho protein to nhn (lm t bo nh l li kt tinh) trong VSV bnh thng b loi b, v khi gy nhim cc VSV ny cho cy n qu, cc vi khun bin i gen c th thay th cc chng bnh thng, nh vy s bo v cy qu khi tc dng gy hi ca sng gi. Nh k thut di truyn, tnh khng nhit v chu hn ca cy trng c ci thin, cng nh c th to ra cc cy trng c tnh khng cn trng v cc VSV gy bnh. Vic to ra v kt hp thnh cng cc vi khun c nh nit c kh nng cng sinh vi cc cy ng cc s lm tng ng k sn lng nng nghip, c bit nhng vng thiu phn bn ho hc. K thut di truyn cng c th cung cp cc bin php mi cho bo v mi trng. Chng hn, mt s vi khun c lai ghp gen phn gii du ho nhm gii quyt cc v trn du ra bin. Cng nghip x l, gii quyt nhim, cng nh cng nghip m v thu hi du cng cn s ng gp ca k ngh di truyn. Tuy nhin k ngh di truyn cng c th gy nhiu tc hi nu nh khng c s kim tra cht ch. Chng hn, vic to dng cc plasmid mi, nu khng c iu chnh mt cch thn trng c th a gen to thnh c t vo cc vi khun m trc khng tn ti. Cc th nghim lin kt ton b hay mt phn ADN t cc virus sinh khi u, hay cc virus khc vo trong cc nhn t ADN sao chp c lp (cc plasmid ca vi khun hay cc ADN virus khc) cng t ra nhng ri ro tng t.
8. DC HC THI HU GENOM NGI

Nm 2004, cc nh khoa hc cng b gii m c b gen ngi. Cng trnh gii m genom ngi thnh cng m ra mt k nguyn mi cho cc ngnh khoa hc s sng trong c y dc hc. Ngi ta c th xc nh trnh t gen ring no ca tng ngi v trn c s so snh vi gen trong b genom chung, c th tin on c p ng ca tng c th i vi cc tc ng khc nhau v nh c th phn nhm c cc c th ang kho st. V phng din Dc hc, ngy nay ang dn hnh thnh mn khoa hc mi l Di truyn dc hc hoc Di truyn dc l (Pharmacogenetics), nghin cu xc nh mt s gen c trng ca cc bnh nhn m cn c vo c th d on trc c cc p ng dng thuc ca bnh nhn, gip trnh c phn ng ph hay phn ng phn v thuc, cng nh h tr la chn thuc chnh xc, an ton hn. V d: C th ngi c gen m ho glucose6phosphat dehydrogenase (G6PD) b t bin khi s dng thuc cha naphthalen hoc mt s dn xut khc c th gy v hng cu, dn n hu qu nghim trng, thm
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ch t vong. Nu nh gii c trnh t gen v xc nh c l c th mang gen t bin th thy thuc s ch nh thuc khc, cha c bnh an ton.
CU HI LNG GI
1. Trnh by cc kiu t bin, c ch tc dng ca cc tc nhn t bin i vi cc VSV Procaryota v ng dng. 2. Trnh by ti t hp di truyn v cc con ng chuyn tnh trng di truyn i vi cc VSV Procaryota. 3. Trnh by vai tr ca cc enzym gii hn, cc phng php a plasmid vo t bo vi khun. 4. Trnh by k thut PCR v phng php xc nh trnh t ADN. 5. Trnh by cc kh nng ng dng ca k ngh di truyn.
PHN HAI
Chng 6
NHIM TRNG
1. KHI NIM NHIM TRNG

Nhim trng (infection) l hin tng xm nhp, pht trin, nhn ln ca VSV trong cc m c th, c th khng biu hin r rt v lm sng, hoc c th gy cc tn thng ti ch, c th tr thnh h
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thng khi lan to ti h bch huyt hoc h tun hon. Khi nim nhim trng cn m ch mt bnh nhim trng. Cn phn nh rng, nhng VSV k sinh bnh thng trn mt s b phn ca c th, nhng khng xm nhp vo m th khng gi l nhim trng. Phn ln chng khng gy bnh, nhng khi gp iu kin thun li th gy bnh, l trng hp nhim trng c hi, cn bnh thng chng c th c li cho c th ch, nh cung cp mt s vitamin, l mt s VSV sng hi sinh (commensalism). Mt s VSV gy bnh i khi cng c kh nng hi sinh tm thi.
2. HNH THI NHIM TRNG

Tu theo mc ly nhim m ta chia nhim trng thnh cc hnh thi chnh sau y:
2.1. Bnh nhim trng

Ta ni n bnh nhim trng trong trng hp VSV xm nhim gy ra ri lon c ch iu ho ca c th, lm mt cn bng ni mi, v dn n nhng biu hin lm sng ca cc du hiu ch quan v khch quan nh st, ho, au, sng ty, ni mn,... c gi l triu chng. Bnh nhim trng c th biu hin cp tnh khi cc triu chng ch tn ti trong khong thi gian ngn, bnh nhn hoc l khi hn hoc l cht. Mt s trng hp cp tnh c th chuyn thnh bnh mn tnh, biu hin bng cc triu chng khng d di nhng li ko di. Th bnh nhim trng ny c th do VSV k sinh ni bo gy ra.
2.2. Nhim trng th n

Ngi b nhim trng m khng c biu hin lm sng nhng c th c thay i cng thc mu, v tm thy cc khng th dch th.
2.3. Nhim trng tim tng

VSV c th c tr ti mt c quan, b phn ca c th ri n mt lc no chng c th gy ra nhim trng r rt. S biu hin trong thc t cc hnh thi nhim trng k trn l hm s tng quan gia VSV v c th, ph thuc vo kh nng gy bnh ca VSV v sc khng ca c th. Yu t mi trng cng c nhng nh hng quan trng ti mi tng quan gia ch th v VSV k sinh. Bn thn VSV cng khng trnh khi cc mi tng quan vi cc VSV khc cng tn ti trong c th. Mi quan h ny c th l i khng (antagonism, antibiosis), hoc hip tc (synergism). Kh nng gy bnh ca VSV c biu th bng c lc ca chng.
3. C LC CA VI SINH VT
c lc l mc gy bnh ca VSV. i a s VSV ch c th gy bnh cho mt loi vt c th, tuy nhin, cng c mt s VSV c th gy bnh cho nhiu loi. Mun nh gi c c lc ca mt chng VSV gy bnh no , ngi ta thng tim (ni chung l gy nhim bnh thc nghim: st ln da b lm xc,...) VSV cho ng vt thc nghim c nhy cm bng dy pha long hn dch chng, ri xc nh liu thp nht cn gy ra p ng, thng thng l ng vt cht. Kt qu c biu th bng liu cht ti thiu (Minimal Lethal Dose) MLD. Tuy nhin, trong thc t kh xc nh c MLD tht chnh xc, v p ng c th ca tng ng vt c tim c khc bit, v th m cn xc nh LD50 (median lethal dose) liu cht trung bnh. y l vic xc nh liu m VSV c th gy cht 50% s ng vt th nghim bng phng php nh gi trn th hoc theo x l thng k. Cc phng php tng t cng c p dng xc nh "cng " ca cc c t. c lc ca VSV bao gm nhiu yu t sau y:
3.1. Kh nng bm vo t bo ch
Bm c vo t bo ch l iu kin u tin VSV c th xm nhp c vo t bo ch v gy ra nhim trng. S hp th c hiu ca virus c bit n t lu, nhng i vi vi khun th vn cn nhiu iu phi nghin cu. Tnh bm c hiu c ghi nhn u tin cc vi khun c tr rut gi. Ti y nhu ng rut rt mnh dn p VSV ra ngoi, nhng nh bm c hiu m trong
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rut vn c c s cn bng v chng loi v s lng vi khun.
3.2. Kh nng xm nhp

Xm nhp l yu t quyt nh i vi nhim trng, v nu khng c xm nhp VSV th cng khng c nhim trng. Khi nim nhim trng bao gm c s nh c trn b mt nim mc (vi khun t, vi khun bch hu,...), c s xm nhp vo bn trong t bo (vi khun l, E. coli, cc virus,...), v vo cc m ca c th (ph cu, t cu vng). i vi cc vi khun gy bnh bng ngoi c t th ngay sau khi phn hu b mt cc t bo ch, ngoi c t do chng tit ra s thm su ngay vo trong c th v gy ra cc ri lon bnh l rt nguy him. i vi cc vi khun c c lc yu th chng thng phi xm nhp su vo bn trong t bo ri mi gy bnh bng ni c t hoc bng cc sn phm chuyn ho ca chng. Cn i vi cc virus v Rickettsia th chng gy bnh bng cch nhn ln bn trong t bo ch.
3.3. c t
c t (toxin) l sn phm chuyn ho ca t bo vi khun, c chia lm hai loi: ngoi c t v ni c t. Ngoi c t l cc cht do cc t bo vi khun sng tit ra. Ni c t l thnh phn ca thnh t bo vi khun v ch c gii phng khi cc t bo ny b ly gii. Chi tit s c trnh by trong chng sau.
3.4. Enzym ngoi bo

Trong qu trnh sng, t bo vi khun tit ra bn ngoi mi trng nhiu loi enzym c gi l cc enzym ngoi bo c lin quan nhiu n kh nng gy bnh ca cc vi khun . Mt s v d c minh ho tip theo y. Hyaluronidase: Enzym ny cn c gi l yu t khuch tn v n c kh nng phn hu acid hyaluronic ca cc m lin kt lm cho vi khun c kh nng lan rng. Nhiu loi vi khun G+ ch tit ra enzym ny. Nu dng cc khng th chng li hyaluronidase iu tr bnh hoi th sinh hi do vi khun Cl. perfringens gy ra th vi khun khng cn kh nng lan to rng ra c. Coagulase l enzym c tc dng lm ng huyt tng, thng do t cu vng v mt s vi khun khc bi tit ra. Enzym ny bin i fibrinogen ca mu thnh fibrin. S lng ng ca fibrin xung quanh t bo vi khun lm ngn cn thc bo v tc dng ca cc khng sinh, khng th. Fibrinolysin: Cc enzym ny cn c tn gi l staphylokinase v streptokinase do t cu v lin cu khun sinh ra. Cc enzym ny bin i plasminogen thnh plasmin dn n hin tng lm tan si huyt v do vi khun d lan trn. Hemolysin: y l enzym c tc dng lm tan hng cu. Nhiu vi khun c kh nng bi tit ra nn c coi l mt tiu chun trong vic pht hin vi khun gy bnh.
3.5. c lc ca virus
c lc ca virus l tp hp ca nhiu yu t gip cho chng nhn ln v gy tn hi cho cc t bo b xm nhim. Ngoi yu t bm v xm nhp, c lc ca virus cn bao gm cc yu t sau: Ngn cn s tng hp cc polyme (i phn t) ca t bo ch phc v cho s nhn ln ca virus. Lm thay i tnh thm ca lysosom v c th dn n s gii phng ca nhiu loi enzym. Gn v peplon vo mng t bo, v lm tn hi cc mng ny. Mt s virus tuy khng c v peplon nhng li gn khng nguyn ca chng lm thay i cu trc v chc nng ca mng t bo. Cc tiu th virus ph hu cu trc v chc nng t bo. Gy bin dng nhim sc th. Lm mt kim sot khng nguyn b mt t bo, gy chuyn dng v lon sn t bo (cc virus gy ung th).
3.6. S n trnh cc p ng min dch

S pht trin c tnh tin ho ca VSV to ra cho chng mt s kh nng chng li cc c ch
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bo v ca c th. S n nu ca VSV trong t bo (vi khun lao, vi khun hi,...) lm cho chng c kh nng trnh c cc tc dng ca nhiu loi khng sinh. Mt s virus gn ADN ca chng vo nhim sc th ca t bo ch ri pht trin nhn ln. Mt s vi khun c th tit (to) ra cc yu t ngn cn khng th, chng hn nh t cu vng tit ra protein A c kh nng kt hp vo phn Fc ca phn t khng th. Mt s vi khun nh ph cu, mng no cu, tit protease thu phn cc Ig. Kh nng bin d ca nhiu loi VSV v cu trc khng nguyn (t cu, vi khun dch hch, virus cm, HIV,...) to cho chng kh nng hn ch tc dng ca min dch c hiu, gy kh khn rt ln cho vic nghin cu cc loi vaccin phng bnh.
4. NGUN GC V PHNG THC TRUYN NHIM

VSV c kh nng gy ra cc bnh nhim trng ngi c ngun gc t 3 ngun truyn nhim: Ngi truyn cho ngi: y l ngun truyn nhim quan trng nht. ng vt truyn cho ngi: Ngun truyn nhim ny ngy cng quan trng hn. Cc vt th t nhin: Tng i khng quan trng. Tm quan trng ca ba ngun truyn nhim k trn tu thuc ch yu vo s khc bit v th cm ca cc VSV khc nhau i vi cc m khc nhau trong c th con ngi.
4.1. Ngi truyn cho ngi

Tr s sinh mi c sinh ra mt cch bnh thng u trong trng thi v trng, nhng ch mt thi gian ngn sau tr c nhiu VSV v dn dn hnh thnh c phc h cc khu h vi khun (bacterial flora). Cc vi khun ny c ly truyn t nhng ngi xung quanh v thay i theo nm thng trong sut cuc i. Thng thng, chng l v hi, nhng v mt l do no nu chng ra khi ni c tr bnh thng th li c th gy bnh, chng hn, E. coli l vi khun khi sng cng sinh trong rut ngi th khng gy bnh, nhng khi rut b thng, chng s gy vim mng bng. Vi khun ny cng thng gy ra cc nhim trng ng tit niu. Streptococcus viridans, Haemophylus influenzae v Pneumococcus l nhng vi khun sng bnh thng nim mc ng h hp trn, nhng c th gy ra vim ph qun, ph qun ph vim, vim xoang, vim tai gia,... nu nh nim mc ng h hp trn b tn thng do virus cm, si, cm lnh. Nhng ngi bnh ang trong thi k bnh (incubation period) ca bnh nhim trng no khi mm bnh ang nhn ln m c th nhng cha c nhng biu hin v lm sng l ngun truyn bnh kh nguy him. in hnh l vi cc bnh nhim virus ng h hp nh si, quai b, bi lit th trong vi ba ngy trc khi pht bnh, ly nhim bnh rt cao. Cc bnh nhn ang trong thi k ton pht l nhng ngi lun o thi ra ngoi mi trng lng VSV rt ln. Tu theo tng loi bnh m cc mm bnh c o thi ra khi c th theo nhng con ng khc nhau. VSV gy bnh v th m c mt trong phn, trong nc tiu, trong m v cc ht nc bt cng nh trong m v trong cc dch bi tit t cc vt lot, vt thng. i vi a s cc bnh cp tnh, VSV gy bnh thng b cht hoc ch c th tn ti mt thi gian ngn trong c th ngi bnh. Tuy nhin mt s VSV gy bnh nh Mycobacterium tuberculosis, Treponema pallidum v Neisseria gonorhae li c khuynh hng gy bnh mn tnh, do cc mm bnh ny thng c o thi ra bn ngoi trong thi gian lu hn. Loi virus Herpes simplex c kh nng tn ti dai dng hnh thnh nn th tim tng trong c th, thnh thong li gy ra cho bnh nhn chng vim lot. Khng phi tt c cc bnh nhn b mc bnh nhim trng u l ngun ly nhim cho ngi khc. Chng hn nh khi mm bnh c c lc rt mnh gy ra ci cht nhanh cho ngi bnh th bn thn mm bnh cng chng cn iu kin tn ti v ly truyn tip. Cc bnh nhn trong giai on phc hi cng i khi l ngun truyn nhim. Mt s bnh nhn sau khi khi bnh bch hu hoc vim hng do lin cu, tuy nhin, do vi khun vn tn ti kh lu trong hng ca h nn vn c mm bnh c th ly nhim cho ngi khc. Sau khi mc cc bnh nh thng hn, l, bi lit, bnh nhn vn cn tip tc o thi VSV ra theo phn. Tu theo thi gian
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o thi mm bnh di hay ngn m nhng ngi ny c phn lm hai loi: Nu thi gian o thi mm bnh ch trong thi gian di 1 nm th gi l ngi mang mm bnh tm thi (temporary carriers); cn nu thi gian o thi l trn 1 nm th c gi l nhng ngi mang mm bnh mn tnh (chronic carriers). Mt s i tng khc c kh nng ly truyn qua ng tip xc. Phn ln h l nhng nhn vin y t do thng xuyn tip xc vi cc VSV gy bnh, nhng h li khng h c cc biu hin bnh l no.
4.2. ng vt truyn cho ngi

Mt s bnh c ngun truyn nhim t ng vt. y cng l nhng ngun truyn nhim ng k. Trong sa ca nhng con b sa b bnh c th c nhng mm bnh nh Mycobacterium tuberculosis, Brucella abortus. Vi khun than (Bacillus anthracis) c th cn sng st nhiu nm trong xc cht ca tru, b, bo t ca chng tn ti c kh lu di trong t v thnh ngun ly nhim bnh cho ngi. Ch b mc bnh di khi cn ngi s truyn virus di gy bnh cho ngi. Loi chut mc bnh dch hch cng c th truyn mm bnh nguy him ny sang cho ngi qua trung gian l b cht chut. Ngy nay, cm gia cm gy bnh cho ngi ang l nguy c i vi nhn loi.
4.3. Cc ngun truyn nhim khc

Mt s loi VSV nh Pseudomonas aeruginosa, Proteus v nhiu chng Clostridium c th tn ti lu di trong t, l ni m chng c th hp th ngun dinh dng t cc cht hu c ca xc ng, thc vt cht. Chng cng thng l cc vi khun sng hi sinh trong ng rut ngi v rut ng vt, v cng t m tr li mi trng t qua phn. Ps. aeruginosa v Proteus thng hay gy ra cc nhim trng vt thng, vt bng v nhim trng ng tit niu. Clostridium tetani v Cl. perfringens gy bnh un vn v hoi th sinh hi khi chng c iu kin xm nhp vo cc m mm trong iu kin k kh v cc vt thng su c ln bn t. Cc loi hp b nhim vi khun Cl. botulinum vi khun k kh sinh ngoi c t rt mnh thng hay gy ra cc v ng c thc phm ng hp.
4.4. Phng thc truyn nhim

Cc mm bnh c th nhim truyn vo c th ngi mnh kho i khi theo con ng trc tip, cn ch yu l gin tip qua cc vt dng, dng c, hay cc mi trng truyn nhim trung gian c coi nh l nhng vt mang thch hp. Cc vt mang c th l bi, khng kh, qun o, sch v, chn mn, nc ung, thc phm v thm ch c cc dng c y t khng v trng. Con ng truyn nhim khc l qua cc cn trng ht mu nh mui, b cht, chy rn, ve,... Tm li, VSV thm nhp vo c th c th qua 4 con ng chnh di y: ng tiu ho. ng h hp. ng da v nim mc. Qua ng nhau thai. Mt s loi VSV gy bnh c kh nng xm nhp vo c th theo nhiu con ng, chng hn nh vi khun lao c th ly nhim qua ng h hp, qua n ung v qua tim truyn. a s cc VSV u xm nhp vo c th v gy ra nhim trng mt khi chng vo theo con ng thch hp c trng. V d nh cc Salmonella ch c th gy ra nhim trng ng rut nu nh i qua con ng tiu ho ch khng th gy ra cc vt thng ngoi da. Nhiu khi ng xm nhp li khng hon ton trng hp vi cc c quan biu hin bnh l, chng hn nh virus bi lit vo c th qua hu hng v ng tiu ho nhng b my thn kinh li l ni xy ra nhng thay i bnh l. 4.4.1. Cc nhim trng ng tiu ho Cc nhim trng ng tiu ho thng thng qua cc con ng truyn nhim sau y: Ly nhim trc tip: Vt dng sinh hot thng thng hay b ly nhim bi phn v qua bn tay. Ngay c nhng ngi c coi l sch s th bn tay h cng thng b nhim cc vi khun c trong
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phn mi khi i i tin. Nhng ngi b bnh tiu chy th kh nng nhim bn bn tay li cng nhiu hn. Vi khun E. coli v Shigella l nhng loi vi khun hay ly nhim theo cch ny nht. Ly nhim qua thc phm: Thc phm l ngun ly nhim kh quan trng trong cc nhim trng ng tiu ho. Sa, tht, c ti sng v k c nhng loi thc n c ch bin sn u rt d b nhim cc loi VSV gy bnh. y l cc mi trng thun li cho chng sinh si ny n nn mt khi chng qua khu thc phm m xm nhp vo ng tiu ho th thng gy ra nhng biu hin bnh l. Ly nhim qua nc: Cc ngun nc sinh hot d b nhim bi nhiu loi VSV khc nhau. Cc loi vi khun nh vi khun t, l, thng hn v ang l nhng mm bnh nguy him gy ra nhiu v dch ln t trc n nay Vit Nam. Cung cp nc sch, x l nc ung hp v sinh cho dn c ang l yu cu cp bch ca nc ta v nhiu nc ang pht trin. 4.4.2. Cc nhim trng ng h hp Ly nhim trc tip: Quan trng nht l s ly truyn trc tip qua khng kh. Tuy nhin, nhng n hn hay thng qua nhng dng c nh cc, a, tha,... cng c xem nh l phng cch trc tip ly truyn qua ng h hp kh quan trng. Ly truyn qua khng kh: Thng qua cc ht nc bt nh cc bnh nhn mc cc bnh nh lao, quai b, cm, si, dch hch,... m cc VSV theo m xm nhp vo ngi khe mnh. Trong khng kh cn thng xuyn c nhng ht bi b gi cun t t ln v c th mang theo mt s vi khun hiu kh, vi khun tan mu, t cu khun, lin cu khun v bo t ca nhiu loi nm mc. Vic m bo thng xuyn c mt bu khng kh trong lnh, t b nhim v mt VSV hc l rt quan trng trong vic phng nga cc cn bnh ly lan theo ng h hp. 4.4.3. Ly truyn qua nhau thai Thng thng th nhau thai l hng ro sinh hc rt hu hiu cho vic bo v bo thai khi cc VSV c trong c th ngi m. Tuy nhin, mt s VSV c bit vn c th xm nhp xuyn qua hng ro ny m nhim vo c th thai nhi. Chng hn, khi ngi m b mc bnh giang mai, xon khun Treponema pallidum c th lun lch vt qua hng ro bo v mng nhau gy ra bnh giang mai bm sinh cho tr s sinh. Khi c thai trong 3 thng u, thai ph mc bnh cm hay bnh Rubella th rt c nhiu kh nng bo thai s b mc cc d tt bm sinh. HIV cng l loi virus c kh nng truyn t cc b m sang cho con trong thi k mang thai. 4.4.4. Truyn qua da v nim mc Bnh thng th da v nim mc ng mt vai tr nh bc tng thnh sinh hc ngn cch gia c th (ni mi) v ngoi cnh. Tuy nhin, trong nhiu trng hp khc nhau, VSV vn c th qua ng da v nim mc m xm nhp vo c th. Do tip xc trc tip: Mt s vi khun, nm mc, k sinh trng c th gy ra cc nhim trng ngoi da ngay c khi da trng thi bnh thng thng qua tip xc hoc l trc tip vi bnh nhn, hoc l thng qua cc vt dng nh qun o, chn mn, khn mi xoa, khn tm,... gy ra cc bnh nh trc u, mn nht, u inh, gh cc, mt s loi nm gy bnh ngoi da, u trng giun mc,... Bnh hi do Mycobacterium leprae gy ra ch yu ly lan qua ng nim mc mi do s dng chung chu ra, khn mt v cc vt dng khc vi bnh nhn. Nhiu bnh v mt nh bnh mt ht, vim kt gic mc cng u ly truyn theo phng cch ny. c bit, mt s VSV c kh nng xm nhp do tip xc trc tip qua nim mc ng sinh dc nh lu, giang mai, h cam, HIV,... c gi l nhng cn bnh ly lan theo ng tnh dc. Ly lan qua cc vt thng: Cc vt thng ngoi da v nim mc chnh l nhng ca ng lm thng thng ni mi v ngoi mi, to iu kin cho rt nhiu cn bnh ly lan theo con ng ny. Vt thng ngoi da v nim mc c th do tai nn, bng, sinh hay do phu thut l nhng nhim trng hay gp nht. Qua cc vt thng ny m nhiu loi VSV xm nhp vo c th gy ra nhiu trng hp bnh l khc nhau. Cc loi vi khun nh S. aureus, S. pyogenes, Cl. perfringens, Cl. tetani l nhng loi vi khun thng gp trong cc nhim trng ly lan qua vt thng. B. anthracis, Leptospira thng xm nhp vo c th ly qua cc vt t tay, chn.
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Ly lan qua con ng tim chch: nhiu loi VSV c c hi xm nhp vo c th qua con ng tim chch hoc qua cc vt t ca nhiu loi cn trng ht mu. Bnh vim gan do virus B gy ra thng ly nhim vo c th qua cc mi tim, qua dng c y t nh km nh rng, cc ng thng, cc dng c ni soi,... Cc loi xon khun nh giang mai, hay k sinh trng st rt cng c th ly lan t ngi ny sang ngi khc qua con ng truyn mu. Cc loi kim tim mi trng bnh vin nu tit trng khng k cng c th l ngun ly nhim HIV, trc khun m xanh, vi khun... Cc loi cn trng ht mu ngi v mt s ng vt l mt trong s nhng tc nhn truyn bnh nguy him. y l nhng vector truyn nhiu loi VSV gy bnh t bnh nhn sang ngi lnh, t ng vt sang ngi. Mui l mt loi cn trng c kh nng truyn nhiu loi bnh nh bnh st xut huyt (st Dengue), bnh vim no Nht Bn B, bnh st rt, bnh giun ch. Cc loi b cht c th truyn vi khun gy bnh dch hch, Rickettsia gy bnh st pht ban dch t. Chy rn truyn cc VSV gy bnh st hi quy, bnh st pht ban dch t. Ve truyn bnh st pht ban do ve, st hi quy Chu Phi, vim no xun h Nga, v nhiu bnh vim no do virus Arbo gy ra. Rui Tse (Tse flies) truyn bnh ng do Trypanosoma rhodesiene gy ra.
5. NHIM TRNG DO THUC

Thuc l cc ch phm dc dng c s dng vi cc mc ch iu tr, phng bnh v chn on, c a vo c th theo nhiu con ng khc nhau. Trong thnh phn ca thuc, ngoi cc hot cht chnh c tc dng dc l c gi l dc cht, th cn c nhiu cht ph gia khc bao gm cc t dc c thnh phn rt khc nhau m trong s c th c cht tr thnh ngun dinh dng cho nhiu loi VSV. Chnh v th m cc loi thuc khc nhau cng thng c th b ly nhim v c th gy ra nhim trng ti cc c s y t v cng ng. Vic thc hin nghim ngt quy trnh sn xut thuc tt theo tiu chun GMP (Good Manufacturing Practice) l mt trong nhng yu cu c t ra tiu chun ho sn xut, hn ch ti a vic ly nhim VSV vo cc thnh phm c sn xut ti cc x nghip dc phm. Khu kim nh vi sinh, khu ng gi bo qun v phn phi thuc n tay ngi bnh cn tun th y cc quy nh hn ch cc s c nhim trng do thuc c th xy ra.
5.1. Nguy c ca thuc b nhim VSV

Thuc b nhim VSV c th gy ra hu qu l lm hng thuc v to ra nguy c cho sc kho ngi dng thuc. Trong nhiu nm theo di ngi ta xc nh c rng, nhiu loi thuc c sn xut ra b hng do nguyn nhn b nhim VSV. Thuc hng c th nhn bit c qua nhng thay i v cm quan, vt l, ho hc, sinh hc. Nhng thnh phn hot tnh c th b VSV chuyn ho lm gim st hiu lc, hoc mt tc dng dc l. Nhng thuc b VSV lm h hng, nu nh vn c s dng iu tr, s gy ra nhng hu qu ng tic cho bnh nhn, nh hng n uy tn v gy ra tn tht v kinh t cho nh sn xut. Bng di y lit k nhng trng hp thuc b nhim VSV t u th k trc n nay.
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5.2. Cc ngun ly nhim

C nhiu ngun ly nhim cho cc sn phm thuc, tuy nhin ch yu l do khu sn xut v s dng. 5.2.1. Ly nhim trong khu sn xut Ly nhim VSV c th xy ra trong khu sn xut hoc trong cc x nghip dc phm hoc trong cc phng bo ch ti cc bnh vin. S nhim khun c th bt u t nguyn liu, nht l cc nguyn liu c ngun gc ph tng ng vt, ri n nc. Cc loi my mc, trang thit b, cc loi thng cha dng trong sn xut nc cng nhiu khi b nhim khun. Khng kh trong nh xng nu khng lc v trng tt l ngun truyn nhim vo cc sn phm. Vt liu dng ng gi, nu khng m bo v trng cng c th l ngun ly nhim vo thuc. V nhn t ngi lao ng tham gia vo dy chuyn sn xut cng c th l ngun ly nhim qua qun o, chn tay,... Cc thnh phm t tiu chun v khun cao chnh l cc ch phm thuc c sn xut trong cc c s sn xut t GMP. 5.2.2. Ly nhim trong khu s dng Thuc cng c th b nhim trng trong qu trnh s dng. Cc loi thuc thng hay b ly nhim VSV nhiu nht l cc dng thuc c ng gi ln dng chung cho nhiu bnh nhn, chng hn nh cc dung mi pha thuc tim, cc loi dch truyn, cc loi thuc dng ngoi, cc loi thuc nh mt (m rng ra l c cc loi m phm). Cc con ng truyn nhim c th l trc tip nh bnh nhn lm cho thuc b nhim, sau , cc thuc b ly nhim ny li truyn sang cho cc bnh nhn khc, hoc theo phng thc cho nh mt bnh nhn no lm cho thuc b nhim VSV ri thng qua m truyn qua bn tay ca cc nhn vin y t. Thng th cc loi vi khun sng cng sinh ngoi da nh Staphylococcus, Micrococcus v cc vi khun Diphteroids l nhng vi khun hay gy nhim cho thuc qua khu s dng.
5.3. Tc hi ca thuc nhim VSV

Tc hi ca thuc b nhim VSV i vi bnh nhn tu thuc vo cc chng VSV xm nhim. Cc VSV ny c phn ra lm hai loi chnh: mm bnh thc s v mm bnh c hi. Khi thuc b nhim cc vi khun nh Clostridium tetani v Salmonella ssp. th c th gy ra tc hi cc k nguy him. Thc t xy ra nhiu trng hp tr s sinh b cht do bnh un vn v xoa phi phn rm m bt talc b nhim Clostridium tettani. Cc ri lon tiu ho do ng c vi khun Salmonella khi ung phi cc ni tit t tuyn gip hoc men tiu ho c iu ch t tuyn tu.
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Cc loi vi khun khc nh Ps. aeruginosa, Klebsiella, Serratia v cc loi vi khun sng t do khc c xem nh l cc mm bnh c hi (opportunist pathogens). i vi nhng ngi kho mnh chng khng gy tc hi g, nhng i vi nhng i tng c bit nh nhng ngi cao tui, nhng bnh nhn b bng, b chn thng hoc nhng ngi b suy gim min dch th chng c th gy ra cc trng thi bnh l nghim trng.
5.4. Cc bin php phng nga v kim tra s ly nhim

Cc bin php phng nga s ly nhim VSV vo cc ch phm thuc bao gi cng tt hn l x l ly nhim. Trong khu sn xut, cc nguyn l thc hnh sn xut thuc tt (GMP) cn c chp hnh nghim chnh; cc thnh phm sau khi sn xut phi c bo qun v ng gi trong nhng thng cha thch hp gim thiu s ly nhim c th xy ra; khu kim nghim nhim khun l yu cu bt buc i vi hu ht cc loi thuc, nht l thuc tim. Vic phng trnh nhim VSV vo thuc trong khu s dng c nhiu kh khn v hn ch. i vi mi trng bnh vin, cc loi thuc ban u c ng lm nhiu liu cn c phn thnh tng liu n cho mi ln s dng trc khi n tay bnh nhn. iu ny rt kh thc hin i vi cc loi thuc dng ngoi v cc dung dch dng pha tim. Tng i d thc hin hn l cc loi thuc vin dng ung. Cui cng bin php bin php hu hiu nht phng trnh ly nhim thuc trong mi trng bnh vin l hun luyn v gio dc cho ngi bnh v cc y t s dng mt cch ti u nht v phng nga tch cc.
6. NHIM TRNG BNH VIN
6.1. Khi nim

Nhim trng bnh vin (Nosocomical infection) l nhng cn bnh b ly nhim trong mi trng bnh vin sau khi nhp vin 48 gi. i vi bnh nhn ang nm vin iu tr bt k loi bnh g cng u c th b ly nhim vi khun, virus, nm,... gy bnh c trong mi trng bnh vin v tr thnh b mc mt hay nhiu loi bnh khc vi cn bnh ban u. Ni rng ra, nhim trng bnh vin cn ch nhng i tng khc nh ngi nh bnh nhn, cc y bc s, y t, iu dng vin, h l v nhn vin cc phng th nghim. Vit Nam cha c thng k chnh thc v nhim trng bnh vin, cn i vi mt s nc pht trin nh Australia trong nm 1979 c t l nhim trng bnh vin l 3,5%, M l 5,2%.
6.2. Ngun truyn nhim

Ngun gc gy ra nhim trng bnh vin phn ln l t mi trng bn ngoi thng qua cc dng c y t b nhim trng, qun o, chn mn ca bnh vin, ngi nh ca bnh nhn v k c i ng nhn vin y t. Rt nhiu loi VSV c th gy ra nhim trng bnh vin, ch yu l trc khun E. coli v cc Proteus. Chim t l cao nht trong cc nhim trng bnh vin l sau cc trng hp phu thut v cc th thut y khoa nh tim truyn, chc d, ni soi,... C quan thng b nhim trng nhiu nht l ng tit niu ri n cc vt m, tip theo l ng h hp, ngoi da, v nhim trng huyt. E. coli l loi vi khun hay gp nht trong nhim trng ng tit niu; Klebsiella trong nhim trng ng h hp; S. aureus trong cc nhim trng ngoi da. Mt khc, Proteus cng rt thng hay gp trong cc ng tit niu, trong cc vt m v nhng nhim trng bnh nhn cao tui. Gn y ngi ta thng cp nhiu n nhng kh nng nhim trng bnh vin gy ra bi trc khun lao, bi cc virus nh HBV, HCV, HIV, v nhiu loi nm nh C. albicans. Bnh nhn, ngi thn v i ng thy thuc l nhng i tng c nguy c cao i vi cc nhim trng loi ny. Nhng VSV gy ra nhim trng bnh vin c th xm nhp vo c th theo tt c cc con ng truyn thng, thng thng, ng tim truyn, ng qua vt m, vt chn thng (tai nn, bng, dng c , no thai, nh rng, ni soi,...), qua tay v qua khng kh l nhng con ng ph bin nht.
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6.3. Cc i tng thng b nhim trng bnh vin

i a s nhng i tng thng b nhim trng bnh vin u l nhng ngi t nhiu c suy gim kh nng khng ca c th min dch. Nhng ngi b mc cc bnh ca c quan min dch, chng hn nh hi chng suy gim min dch bm sinh hay mc phi (HIV/AIDS). Nhng ngi dng thuc p ch min dch (immunosupressors). Nhng bnh nhn sau khi phu thut, b bng nng, bnh nhn b mc cc bnh v ni tit nh i tho ng, bnh thiu nng tuyn gip, nhng bnh nhn gi c, tr nh v nhng tr b suy dinh dng hay nhng ngi c iu tr khng sinh lu di.
6.4. Cc nguyn tc phng nga

Phng nga bao gi cng tt hn so vi x l i vi cc nhim trng bnh vin. Cc nguyn tc chnh cn tun th: Ci thin v tng cng sc khng ca bnh nhn bng ch bi dng, n ngh v iu tr thch hp. Lm trong sch, ci thin mi trng bnh vin, thng xuyn ty u v tit trng phng m, phng pha ch v bnh phng. Thc hin x l nghim ngt tt c cc cht thi ca bnh nhn c gi chung l cc cht thi bnh vin. Tit trng tt, m bo v sinh cc dng c y t, qun o, ging chiu, chn mn,... v gio dc thc cho i ng nhn vin y t v ngi nh bnh nhn. Trong khi iu tr, nht l i vi iu tr s dng cc loi khng sinh cn tun th ng cc quy nh iu tr, hn ch ti a s nhn thuc dn n vic hnh thnh cc chng vi khun khng khng sinh t trong bnh vin, ri t ly lan ra cng ng x hi.
CU HI LNG GI
1. Trnh by khi nim nhim trng, cc hnh thi nhim trng; trnh by cc yu t to c lc ca VSV gy bnh. 2. Trnh by ngun gc v phng thc truyn nhim; trnh by tc hi ca thuc b nhim VSV.
Chng 7
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1. KHI NIM V P NG MIN DCH

Min dch (immunity) l kh nng c th nhn ra v loi b cc vt l. p ng min dch chia hai loi: min dch t nhin v min dch thu c
1.1. Min dch t nhin (Native immunity) Min dch khng c hiu
Min dch t nhin (hay min dch khng c hiu) l kh nng t bo v sn c v mang tnh di truyn trong cc c th cng mt loi. Kh nng ny c ngay t lc mi sinh v khng cn phi c s tip xc trc ca c th vi cc yu t l. Min dch t nhin ng vai tr l tuyn phng th u tin ngn chn s xm nhp v tiu dit VSV trc khi chng kp nhn ln trong c th, nh m h min dch c hiu c thi gian hnh thnh. Gi l min dch khng c hiu l do n p ng min dch vi tt c cc loi khng nguyn l nh nhau. 1.1.1. Cc hng ro ca p ng min dch t nhin 1.1.1.1. Hng ro vt l Da, nim mc ngn cch mi trng trong c th vi mi trng bn ngoi. Da bao ph mt ngoi c th, c cu to gm lp biu b cha cc t bo biu m sp xp ken cht to hng ro c hc ngn cn s xm nhp ca VSV. Trn mt lp biu b l lp t bo sng ho cha keratin, khng thm nc, cc VSV khng phn gii c keratin s khng th theo nc ngm vo c th. Nim mc bao ph mt trong c th nh ng tiu ho, h hp, tit niu, sinh dc... Nim mc ch c mt lp t bo trn c ph bi lp cht nhy, to mt mng bo v ngn khng cho cc VSV l bm vo lp t bo m hoc c quan. Trn b mt nim mc h hp c nhiu lng mao chuyn ng lin tc theo mt hng cn cc vi sinh t bi v y chng ra ngoi bng phn x ho, ht hi. B mt nim mc (mt, ming, tit niu) lun tit dch (nc mt, nc bt, nc tiu) to ra dng dch th ra tri VSV trn b mt. 1.1.1.2. Hng ro ho hc Cc VSV nu qua c hng ro vt l th vp phi hng ro ho hc l: acid trn mt da nh c cht tit to toan nh acid lactic, acid bo ca m hi, tuyn m di da m cc VSV khng tn ti lu c. Dch v do d dy tit ra c pH 1 2 nn a s VSV xm nhp vo bng ng tiu ho khng sng st c. Lysozym: Dch tit ca cc tuyn nh nc mt, nc bt, nc mi, sa c cha lysozym, l mt cht c kh nng c ch s tng hp thnh t bo vi khun Gram dng. Cc protein gn st l nguyn t cn thit cho sinh trng ca VSV. Mt s protein gn st
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nh lactoferin, transferin lm gim nng st t do trong mu xung rt nhiu ln so vi nhu cu ca VSV. 1.1.1.3. Hng ro th dch Cc yu t bo v c trong mu v dch ni mi nh: Interferon (IFN) l nhm glycoprotein cm ng, c sn xut bi nhiu loi t bo nh t bo bch cu, i thc bo, t bo biu m, t bo s non khi c cm ng bi virus hoc acid nucleic. IFN c hot tnh khng virus mt cch khng c hiu nh ngn cn s nhn ln ca virus, hot ho cc t bo dit t nhin. IFN c kh nng chng li cc t bo ung th do c ch t bo trng thnh, c ch s phn bo. B th l h thng nhiu protein thnh phn, c hot ho theo mt trnh t nht nh. Hot ng ca b th gy tn thng thnh t bo, sau gy tan bo, ng thi gip tng cng hin tng thc bo. 1.1.1.4. Hng ro t bo Trn nim mc c rt nhiu t bo c kh nng thc bo. Chng c Metchnikoff pht hin t u th k XX, gm hai loi: tiu thc bo (microphage l nhng bch cu a nhn trung tnh ca mu), i thc bo (macrophage l nhng t bo bt ngun t tu xng, phn ho thnh mono bo mu ri di tn ti cc m). Qu trnh thc bo chia ba giai on: Giai on gn: Cc VSV khi gp cc t bo thc bo s b dnh vo mng t bo nh cc receptor b mt khc nhau. Giai on nut: Mng t bo lm vo, cht nguyn sinh s to ra cc chn gi bao ly VSV, ri ng kn li to hc thc bo Phagosom. Giai on tiu: Cc ht lysosom tin n gn hc thc bo v xy ra hin tng ho mng, mng lysosom nhp vo cng mng phagosom gi l phagolysosom. Cc cht c trong lysosom s vo trong hc thc bo tiu dit i tng cn thc bo. Trong phagolysosom cc VSV s b tiu dit bng: pH acid, enzym thu phn, superoxyt.... T bo dit t nhin (Natural killer NK) l t bo dng lympho c ht ln trong sinh cht, c mu ngoi vi, c chc nng dit cc t bo ch (t bo b nhim virus, t bo ung th) bng cht tit (perforin). Hot tnh ny ca NK tng ln khi c kch thch bi interferon. T bo K (Killer cell) l t bo dng lympho, cn gi l t bo null, trn b mt t bo c th th dnh cho Fc ca khng th, chc nng gy c t bo ch nh hiu qu ADCC (Antibody dependent cellular cytotoxicity gy c t bo ph thuc khng th). 1.1.1.5. Hng ro VSV Mc d b mt c th c bo v chng li s xm nhp ca VSV nhng mt s loi nht nh vn c kh nng sng, l h khun ch bnh thng ca c th. a s chng l vi khun trong ng tiu ho, mt s phn b t nhin v to qun th trn da, xoang ming, ng h hp, ng sinh dc,... Thng thng, cc VSV ny khng gy bnh, chng ch pht trin trn b mt m khng xm nhp su vo bn trong m, chng cnh tranh v tr bm, thc n, lm gim nng oxy, tit mt s cht dit khun gy bt li cho cc VSV xm nhp gy bnh. H khun ch bnh thng l mt qun th phc tp v cn bng. Nu trng thi cn bng b mt, cc VSV khng gy bnh tr thnh gy bnh, gi l nhng VSV gy bnh c hi. 1.1.1.6. St Thn nhit ngi lun n nh mc 37OC. St l s tng thn nhit, l c ch bo v t nhin ca c th, st lm tng tc phn ng enzym phn hu VSV, tng hot ng ca interferon, gim nng st t do trong mu. 1.1.1.7. Vim khng c hiu Phn ng vim c to thnh nhm khu tr cc VSV mi xm nhp vo mt ni, khng cho chng lan rng v tiu dit chng. Hin tng ny c nu vi 4 triu chng kinh in l: sng,
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nng, , au. Sng l do tng thm thnh mch, tit dch tp trung quanh t bo m. Tng tnh thm ko theo s xuyn mch ca cc t bo thc bo ti vim. Nng l hu qu ca s dn mch, dng mu tng cng ti ni nhim trng lm nhit tng ln. do cc t bo bch cu kim v t bo mast tit cht hot mch, gy dn mch, dng mu dn n nhiu hn. au trong trng hp vim l do t bo tit ra bradykinin v prostaglandin kch thch dy thn kinh gy au. Nh vy, v c bn, phn ng vim l c li, n ch gy hi khi phn ng qu mc, gy nhim toan, ri lon thng bng nc in gii... Tm li, khi mt VSV l xm nhp vo c th, chng s gp phi hng lot cc c ch bo v khng c hiu trong p ng min dch t nhin vi mc ch tiu dit v loi tr chng ra khi c th.
1.2. Min dch thu c hay Min dch c hiu (specific immunity)
1.2.1. Khi nim Min dch thu c hay min dch c hiu l trng thi min dch xut hin khi c th c tip xc vi khng nguyn (a vo ch ng hay tip xc ngu nhin). Min dch c hiu c th c khi c truyn khng th hoc cc t bo min dch. p ng min dch c hiu c 2 hnh thc l: Min dch dch th (Humoral immunoresponse) v min dch qua trung gian t bo (Cell mediated immunorespose). 1.2.2. c im c bn ca p ng min dch c hiu l: Tnh c hiu. Phn bit l quen. Tnh a dng. Tr nh min dch. iu ho min dch. 1.2.3. Vim c hiu Phn ng vim c hiu xy ra khi c th tip xc vi khng nguyn v c khng th dch th hoc khng th t bo. S kt hp khng nguyn vi khng th c hiu khi pht phn ng vim cng gm 4 triu chng: sng, nng, , au. 1.2.4. Phn loi min dch c hiu 1.2.4.1. Min dch ch ng (active immunity) L trng thi min dch ca mt c th do h min dch ca bn thn c th sinh ra khi c khng nguyn kch thch. Min dch ch ng chia lm 2 loi: Min dch ch ng t nhin l trng thi min dch c c khi c th v tnh tip xc vi khng nguyn. Min dch ch ng thu c l trng thi min dch c c khi khng nguyn c ch ng a vo c th, nh tim vaccin. 1.2.4.2. Min dch th ng (passive immunity) L trng thi min dch c c nh khng th c truyn t ngoi vo. Min dch th ng chia lm 2 loi: Min dch th ng t nhin l trng thi min dch c c do khng th c truyn t m sang con qua rau thai hoc qua sa. Min dch th ng thu c l trng thi min dch c c khi khng th c a vo c th, nh dng liu php huyt thanh.
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S khng ca c th gm 2 hnh thc: p ng min dch t nhin v p ng min dch c hiu. Hai hnh thc ny lun b sung, h tr v khng th tch ri nhau. Min dch t nhin c trc ngn cn VSV trc khi chng kp nhn ln trong c th v nh m h thng min dch c hiu c thi gian hnh thnh v ng vai tr ch cht.
2. CC C QUAN V T BO THAM GIA VO QU TRNH MIN DCH

p ng min dch l mt qu trnh bo v rt quan trng v phc tp, l kt qu ca s hp tc nhiu loi c quan, t chc v t bo khc nhau nhn din v phn ng vi khng nguyn.
2.1. C quan lympho

C quan lympho l ni sn sinh, hun luyn v tng tr cc t bo lympho, c chia thnh: c quan lympho trung ng v c quan lympho ngoi vi. 2.1.1. C quan lympho trung ng L ni bit ho t bo ngun thnh t bo lympho chn khng cn s kch thch ca khng nguyn. 2.1.1.1. Tuyn c (Thymus) l mt khi dt gm 2 thu nm st ngay sau xng c, y l c quan lympho xut hin sm nht trong thi k bo thai, n pht trin v kch thc trong thi k ny v sau khi tr ra i. n tui dy th l pht trin ti a (30 50g) v thoi trin dn v ch cn di 15g ngi gi. Tuyn c c cu to bi cc t bo dng lympho v cc t bo dng biu m. Mi thu tuyn c li chia thnh nhiu tiu thu. Mi tiu thu c chia 2 vng: vng v v vng tu.
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Cc t bo ngun t tu xng di c n tuyn c, ti vng v cc tin t bo T c bit ho phn chia nhiu ln thnh t bo T chn v i vo vng tu. Vng tu l ni tip tc bit ho ca cc t bo T chn thnh cc lympho bo T chc nng v ri tuyn vo mu n cc c quan lympho ngoi vi. Nh vy, tuyn c l ni m nhn chc nng hun luyn, phn chia, bit ho cc lympho bo dng T. 2.1.1.2. Bursa Fabricius (ti Fabricius) Ti Fabricius ch c loi chim, loi g, l c quan lympho nm st nhp. Ti cha cc nang lympho v l ni bit ho t bo ngun thnh t bo lympho B chn. ng vt c v, t bo ngun c bit ho thnh t bo lympho B chn ngay ti tu xng.
Cc t bo lympho B chn vo mu v n c quan lympho ngoi vi, chng tip xc vi khng nguyn, bit ho tr thnh t bo plasma sn xut khng th dch th. 2.1.2. C quan lympho ngoi vi L ni c tr ca cc t bo lympho c bit ho. 2.1.2.1. Lch l mt c quan lympho ln c chc nng: lc, d tr mu cho c th, tp trung khng nguyn xm nhp bng ng tnh mch ri to p ng min dch chng li.
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Nhu m lch c chia lm 2 phn: tu chim 4/5 khi lng lch v tu trng l nhng im ri rc xen vo khi tu . Tu trng c hai vng: mt vng c cc nang lympho cha cc tm im mm ca dng lympho B, gi l vng khng ph thuc tuyn c, mt vng cha cc lympho bo T gi l vng ph thuc tuyn c. Tu c vai tr nh mt ci lc i vi hng cu b hu hoi do tn thng hoc do gi, cc mnh t bo cht. Tu c nhiu xoang tnh mch cha hng cu, bch cu, tiu cu v lympho bo. 2.1.2.2. Hch lympho Hch lympho cn gi l hch bch huyt c hnh ht u c bc trong mt v lin kt. Hch nm ri rc trn ng i ca mch bch huyt, v thng tp trung thnh m ti ch giao nhau ca mch bch huyt nh c, nch, bn... Hch c ng knh khong 1 25mm, chng to ln r rt khi b nhim khun, b u c tnh. Dch bch huyt thu thp t mt vng nht nh trong c th c dn vo hch, sau ra khi hch ti hch khc hoc chy vo ng ngc (c quan bch huyt ln ca c th). Hch gm cc thu, mi thu chia hai vng: vng v v vng tu. Vng v li chia hai: Vng v nng c nhiu nang lympho cha t bo lympho B, cn gi l vng khng ph thuc tuyn c. Vng v su cha ch yu l t bo lympho T, ngoi ra, cn c i thc bo v mt t t bo lympho B c gi l vng ph thuc tuyn c. Vng tu l trung tm hch c nhiu lympho T, i thc bo, t y cc t bo ri hch ra ngoi. Hch hot ng nh mt ci lc vi cc phn t l, l ni cc t bo min dch tip xc vi khng nguyn. Cc vt l phi di chuyn theo nhng mch nh v hp vi vn tc nh tip xc vi i thc bo v cc t bo lympho. i khi VSV vt qua hch trc nhng li b gi li hch sau. Khi b nhim trng, VSV vt qua tt c hch vo mu, tuy nhin, h thng hch lm chm li s nhim trng kp thi huy ng h min dch c hiu.
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2.1.2.3. Cc c quan lympho khc Cc m lympho rut: Gm mng Peyer v cc nang lympho nm ri rc nim mc rut. Mng Peyer l nhng nang lympho m trung tm cha nhiu lympho B, lympho T v i thc bo. Cc t bo lympho s tip xc vi khng nguyn xm nhp qua ng tiu ho, c vai tr quan trng trong c ch bo v ti ch. Hch hnh nhn l nhng m lympho hng. Cc hch hnh nhn hp thnh vng Waldeyer: hch nhn li, khu ci, hu, vi. mt s ni khc nh: ph nang, ph qun, ng niu, ng sinh dc cng cha m lympho nm di lp nim mc.
2.2. Cc t bo tham gia p ng min dch

Cc t bo ca h thng min dch u c sinh ra t cc t bo ngun tu xng. Trong qu trnh bit ho to ra dng tu, dng hng cu v dng lympho.
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Dng to mu tip tc bit ho thnh ba dng: dng hng cu, dng t bo to tiu cu v dng tu. Cc t bo dng tu li bit ho thnh hai nhnh: mt nhnh to t bo n nhn ri thnh i thc bo, mt nhnh to t bo a nhn ri phn ho thnh bch cu trung tnh, bch cu a kim, bch cu a acid. Dng t bo lympho vo c quan lympho trung tm bit ho thnh cc t bo chn. T bo NK (Natural killer), t bo K (killer) l nhng lympho bo khng bit ho trong c quan lympho trung tm. 2.2.1. T bo lympho T bo lympho c trong tun hon mu v bch huyt, c hai loi t bo lympho: lympho T v lympho B. 2.2.1.1. Lympho T Lympho T l t bo chu trch nhim p ng min dch qua trung gian t bo. Lympho T chim 70% tng s lympho bo mu ngoi vi v chim a s cc m lympho. T t bo ngun tu xng, lympho T c bit ho v trng thnh tuyn c (Thymus, nn k hiu l T). Qu trnh bit ho t bo T tuyn c to ra hai dng t bo T: mt dng cha khng nguyn
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T4 (hay CD4) gi l t bo T4, mt dng cha khng nguyn T8 (hay CD8) gi l t bo T8. Dng t bo T4 li tip tc bit ho to thnh cc phn lp: + Lympho T h tr TH (Hepper T cells): nhn bit khng nguyn, thc y hot ng ca cc lympho T khc v lympho B. + Lympho T gy qu mn mun TDTH (Delayed type hypersensitivity T cell) c chc nng hot ho i thc bo v bch cu gy qu mn mun. + Lympho T iu ho ngc TFR (Feedback regulator T lymphocyte): hot ho lympho T c ch. Dng T8 tip tc bit ho thnh cc phn lp: + Lympho T c t bo TC (Cytotoxic T lymphocyte): nhn bit khng nguyn, tn cng trc tip cc t bo c khng nguyn l trn b mt (t bo ch). + Lympho T c ch TS (Suppessor T lymphocyte): c nhim v iu ho min dch, c ch hot ng cc lympho khc. Cc dng t bo T ny ri tuyn c vo tun hon mu to qun th t bo T cc c quan lympho ngoi vi. 2.2.1.2. T bo lympho B Lympho B l t bo chu trch nhim p ng min dch dch th. Qu trnh bit ho t bo B ca chim, gia cm ti Bursa Fabricius, ngi l tu xng (Bone marrow) khng cn s kch thch khng nguyn. T bo ngun tin B B cha chn B chn. Lympho B chn ngi c c trng bi s hin din ca th th globulin b mt (SIg: Surface Immuoglobulin). Cc lympho bo B vi cc SIg b mt n cc m lympho ngoi vi, khi c s kch thch ca khng nguyn th li bit ho, tng sinh thnh cc tng bo sn xut khng th. B chn B bit ho, tng sinh tng bo khng th dch th. 2.2.1.3. Di chuyn v ti tun hon ca cc lympho bo Sau khi trng thnh tu xng, tuyn c, cc lympho bo theo h tun hon di chuyn n cc m lympho ngoi vi. Nu khng c khng nguyn kch thch th cc lympho bo cht sau 2 3 ngy. Nu c khng nguyn kch thch th sng khong 120 140 ngy, ri t hch i theo ng bch huyt ra ng ngc ri tr li tun hon mu.
2.2.1.4. T bo NK (Natural Killer Cell) NK l nhng t bo lympho trong mu ngoi vi khng phn loi c l t bo T hay B. NK c kh nng dit cc t bo ch nh: t bo u, t bo ch nhim virus.
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2.2.1.5. i thc bo i thc bo l nhng t bo c kch thc ln c kh nng bt gi, x l khng nguyn cng nh hp tc vi cc t bo lympho. i thc bo c hai loi: + Loi c nh hch, lch, tu xng, gan. + Loi di ng trong mu v bch huyt. S tham gia ca i thc bo trong p ng min dch: + Thc bo khng nguyn l xm nhp. + Trnh din thng tin v khng nguyn l cho cc t bo lympho. + C th th vi Fc ca khng th v b th C3b. + Tit mt s cht c hot tnh sinh hc nh: lysozym, interferon,... 2.2.1.6. Bch cu ht trung tnh (BCTT) BCTT cn gi l tiu thc bo, c chc nng bt, tiu dit vi khun. BCTT chim 60 70% tng s bch cu mu ngoi vi, i sng khong 4 5 ngy, c kh nng vn ng mnh bng gi tc, d dng lch qua thnh mch n vim, BCTT c th th vi khng th v C3 b th. 2.2.1.7. Bch cu i kim (BCAK) BCAK ch chim 0,5% tng s bch cu ngoi vi, c vai tr quan trng trong phn ng phn v v d ng do b mt c th th vi IgE, bo tng cha cc ht c hot tnh sinh hc nh: histamin, serotonin, heparin. 2.2.1.8. Bch cu i toan (BCAT) BCAT chim 1 5% tng s bch cu ngoi vi. BCAT tng trong cc trng hp nhim k sinh trng, c ch ca hin tng ny th cha r.
3. KHNG NGUYN
3.1. nh ngha
Khng nguyn (Antigen Ag) l cht c kh nng: Kch thch c th sinh p ng min dch. Kt hp c hiu vi khng th hoc vi cc th th t bo lympho.
3.2. Tnh cht ca khng nguyn

3.2.1. Tnh sinh min dch Tnh sinh min dch ca mt khng nguyn ph thuc: 3.2.1.1. Tnh l ca khng nguyn L s khc bit chng loi gia tc ch v khng nguyn, cht cng l th kh nng kch thch min dch cng mnh. 3.2.1.2. Khi lng phn t Nhng cht c phn t lng ln hn 1.000 dalton mi c kh nng sinh min dch (tr mt s trng hp ngoi l) v khi vo c th chng gn vi nhng cht c phn t lng ln. 3.2.1.3. Cu trc phn t phc tp Protein: phn ln cc protein u c kh nng sinh min dch (tr mt s t nh gelatin). Cc protein rt d b phn hu bi enzym nn nu vo c th bng ng tiu ho th rt d b mt tc dng. Lipid khng c tnh khng nguyn nhng khi gn vi protein (lipoprotein) th tr thnh khng
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nguyn. Polysaccharid: Tnh khng nguyn yu, v cu trc lp li, thiu a dng, khi vo c th chng d b phn gii. Acid nucleic l khng nguyn yu nhng nu gn vi protein (nucleoprotein) th tnh khng nguyn tng. 3.2.2. Tnh c hiu ca khng nguyn Mi khng nguyn c mt cu trc c hiu ring c gi l quyt nh khng nguyn (epitop), y l phn quan trng ca khng nguyn ni cc t bo min dch v khng th nhn dng v gn c hiu. Phn kt hp vi mt epitop khng nguyn trn mi phn t khng th gi l v tr kt hp khng nguyn (paratop), cn trn t bo lympho l th th (receptor). Mi epitop ch gn c hiu vi mt paratop hoc mt th th t bo lympho v ch sinh mt dng khng th c hiu.
3.3. Cc loi khng nguyn

Khng nguyn n gi c mt epitop. Khng nguyn a gi c nhiu epitop khc nhau. Hai khng nguyn cho l hai khng nguyn khc nhau nhng c mt hoc nhiu epitop ging nhau.
3.4. Phn loi khng nguyn

3.4.1. Theo cu trc ho hc Khng nguyn protein. Khng nguyn polysaccharid. Khng nguyn lipit. Khng nguyn acid nucleic. 3.4.2. Khng nguyn VSV 3.4.2.1. Khng nguyn vi khun Ngoi c t l nhng cht c do vi khun tit ra ngoi, bn cht l protein. Ngoi c t c th x l vi formalin 0,5% 37OC trong 1 n 2 thng kh tnh c gi l gii c t (anatoxin), nhng cn tnh khng nguyn v c ng dng iu ch vaccin phng bnh. Enzym ngoi bo c lc mt s vi khun nh hyaluronidase, leucocidin, hemolysin, coagulase,... Cc enzym ny l khng nguyn tt kch thch c th to khng th c hiu. Mt s enzym c s dng trong chn on nh streptolysin O ca lin cu. Khng nguyn vch (khng nguyn O), vi khun Gram(+), cc cht nh polysaccharid, protein vch c tnh khng nguyn v tu loi vi khun m c cc lp trong vch quyt nh tnh c hiu khng nguyn thn. vi khun Gram() c cc lp khng nguyn vch gn nh nhau, l LPS (lipopolysaccharid), tnh c hiu ca khng nguyn O c quyt nh bi lp polysaccharid ngoi cng. Khng nguyn v (khng nguyn K). Cc vi khun c v th v c tnh khng nguyn. Bn cht ho hc l polypeptid hoc polysaccharid nn kh nng sinh min dch khng cao, nhng khi gn vi t bo vi khun vn gy c min dch. Khng nguyn lng (khng nguyn H). Si lng ca vi khun to thnh bi cc protein si (Flagellin). Khng nguyn lng c dng phn loi mt s vi khun.
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3.4.2.2. Khng nguyn virus Cc khng nguyn ho tan l nhng khng nguyn thu c t nui cy t bo nhim virus sau khi loi b virus v cc thnh phn ca t bo. Cc khng nguyn ny c th l enzym, l thnh phn cu to m virus tng hp tha hoc khng nguyn b mt bong ra. Cc khng nguyn cu to virus: + Khng nguyn li virus: Acid nucleic l bn khng nguyn nhng nucleoprotein l khng nguyn hon ton ca virus. + Khng nguyn v envelop: V envelop thng l lipoprotein hoc glycoprotein. Trn v thng c nhng khng nguyn c hiu nh hemaglutinin (khng nguyn H: yu t gy ngng kt hng cu) hoc neuraminidase (khng nguyn N: enzym ph hu im tip nhn trn b mt t bo cm th). Ngoi ra, trn v ca mt s virus cn c cc gai nh gn vo c tc dng lm cho virus bm vo t bo cm th nh HIV. y l nhng khng nguyn quan trng trong chn on HIV.
4. TRNH DIN KHNG NGUYN PHN T MHC

MHC (Major Histocompability Complex) l phc hp ho hp m ch yu c vai tr quan trng trong trnh din khng nguyn. Phn t MHC hay cn gi l khng nguyn MHC, l nhng phn t protein biu l trn b mt nhiu loi t bo trong c th, n khin c th ny khc c th kia nu khng phi l sinh i cng trng hoc khng cng mt dng thun chng. Phn t MHC lp I c trn b mt tt c t bo c nhn ca ng vt c xng sng. Phn t MHC lp II c trn b mt t bo lympho B, i thc bo, v mt s t bo trnh din khng nguyn khc nh t bo tua Deudritic, t bo Langerhans. Cc khng nguyn khng th trnh din trc tip cho t bo lympho B hoc lympho T, m phi c cc t bo trnh din khng nguyn (Antigen Presenting cell APC) x l, ngha l chuyn cc protein phc tp thnh cc on peptid ph hp vi rnh gn peptid trn phn t MHC ca t bo APC . ngi, khng nguyn ph hp t chc l HLA (Human Leucocyte Antigen) khng nguyn b mt bch cu ngi.
4.1. Cu trc ca cc phn t MHC

4.1.1. Phn t MHC lp I Phn t MHC lp I l cc glycoprotein gm 2 chui polypeptid: chui cm su vo mng sinh cht, vng c u tn amin ngoi bo trn chui mang cc rnh gn peptid; chui l mt polypeptid nh khng gn trc tip vi t bo. 4.1.2. Phn t MHC lp II Phn t MHC lp II l phn t protein gm 2 chui polypeptid: chui v chui kt hp khng ng ho tr vi nhau v cng cm su vo mng sinh cht. on amin ngoi bo ca c 2 chui to nn cc rnh gn peptid. Cu trc phn t MHC gia cc loi ng vt, thm ch gia cc c th trong cng mt loi l khng ging nhau. iu ny gii thch hin tng thi loi khi ghp m gia hai c th.
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4.2. S trnh din khng nguyn ngoi bo kt hp vi phn t MHC lp II

T bo trnh din cc khng nguyn ngoi bo thng gp l i thc bo v lympho B, t bo tua Deudritic. Khng nguyn protein l c cc t bo APC tm bt v a vo trong ni bo, sau chng c ct thnh nhng on peptid nh, thng, nhng cn kh nng sinh min dch v c tch t li trong cc nang endosom. Cc phn t MHC lp II c tng hp trong li ni cht v c che ph bng cc protein bao vy (blocking protein protein ny ngn khng cho phn t MHC lp II gn vi cc peptid khc cng c to ra trong li ni cht). Sau phn t MHC lp II cng protein bao vy i vo endosom. Ti y protein bao vy c phn gii, do , cc phn t MHC lp II s c gn vi on peptid khng nguyn to phc hp MHC lp IIpeptid khng nguyn v c chuyn ln b mt t bo APC trnh din cho lympho T. Lympho T n tip xc trc tip vi t bo APC v hnh thnh cp kt dnh CD4MHC lp II v TCRpeptid khng nguyn, nhn din khng nguyn l.
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4.3. S trnh din khng nguyn ni bo kt hp vi phn t MHC lp I

Khng nguyn ni bo l loi khng nguyn c hnh thnh trong t bo ch, c th l cc protein virus, khng nguyn ung th, hoc t khng nguyn. Cc khng nguyn ny c t bo ch x l bng cc enzym phn gii thnh cc peptid. Cc peptid ny gn vi phn t MHC lp I trong li ni cht, to phc hp MHC lp I peptid khng nguyn ri chuyn ln b mt t bo. Lympho T n tip xc trc tip vi t bo v hnh thnh cp kt dnh CD8 MHC lp I v TCRpeptid khng nguyn nhn din v tit cht c tiu dit t bo mang khng nguyn (t bo ch).
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5. P NG MIN DCH DCH TH
5.1. T bo lympho B
T bo lympho B l t bo sinh khng th dch th, c bit ho ti Bursa Fabricius i vi chim v gia cm, ngi l tu xng, sau , cc t bo B chn vo mu ri n c tr ti lch v hch ngoi vi.
5.2. Qu trnh tng sinh, bit ho t bo lympho B

Di knh hin vi in t, cc t bo lympho B c b mt x x, ni gai, l cc globulin min dch b mt SIg (Surface immunoglobuline). Qu trnh tng sinh, bit ho lympho B din ra km theo s thay i SIg. Qu trnh gm 2 giai on: 5.2.1. Giai on 1 Cc t bo ngun trong tu xng pht trin thnh tin lympho B, cc t bo ny cha c SIg. Tip theo, tin lympho B thnh lympho B cha chn ri thnh lympho B chn vi s xut hin ca SIg. Mi lympho B c khong 0,5 1,5. 105 phn t SIg, chng hot ng nh cc th th b mt tip nhn khng nguyn. giai on ny, s pht trin ca lympho B khng cn s kch thch ca khng nguyn v s h tr ca lympho T. 5.2.2. Giai on 2 Giai on ny cn c s kch thch ca khng nguyn v s h tr ca t bo lympho TH. Khng nguyn khi vo c th s chn lc gn vi lympho B chn c SIg thch hp, to phc hp khng nguyn SIg. Lc ny, t bo lympho B s tri qua mt qu trnh tng sinh, bit ho thnh dng t bo plasma (tng bo) sn xut khng th dch th v dng t bo nh (menmory B cell) gip cho p ng ln sau vi chnh khng nguyn nhanh v mnh hn.
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5.3. Khng th dch th (Antibody)

L cc globulin c trong huyt thanh c kh nng lin kt c hiu vi khng nguyn kch thch sinh ra n. Khng th min dch Immunoglobulin k hiu l Ig. Hai c tnh sinh hc quan trng ca khng th l kh nng lin kt c hiu vi khng nguyn v kh nng biu hin nh mt khng nguyn, tc l kch thch sinh khng khng th. 5.3.1. Cu trc Phn t globulin min dch gm mt hay nhiu n v hnh thnh. Mi n v l mt phn t protein gm 4 chui polypeptit ging nhau tng i mt: 2 chui nh v 2 chui nng ni vi nhau bng cu disulfua.
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Hnh 7.12a. Cu trc ca khng th
5.3.1.1. Chui nh, k hiu L (Light chain) Chui nh c khi lng phn t khong 23.000 cha 211 221 acid amin, c 2 loi chui nh chung cho tt c cc lp globulin min dch. Chui nh Kappa (k hiu ). Chui nh Lambda (k hiu ). Mi phn t globulin ch cha mt loi chui nh hoc Kappa, hoc Lambda m khng bao gi cha c hai. Mi chui nh cha hai vng acid amin. Vng c trt t acid amin khng thay i gi l vng c nh, k hiu l C (constant), vng ny nm pha u carboxyl (COOH). Trt t acid amin vng ny lun ging nhau tt c cc lp khng th. Vng c trt t acid amin thay i gi l vng bin i, k hiu l V (variable), vng ny nm pha u amin (NH2). Trt t acid amin vng ny khc nhau k c cc phn t globulin do cng mt t bo sinh ra. 5.3.1.2. Chui nng k hiu H (Heavy chain) Chui nng c khi lng phn t t 50.000 n 70.000, chia thnh 5 loi: , , , , , tng ng vi 5 lp khng th l IgG. IgM, IgA, IgD, IgE. Cc chui nng c tnh c hiu ring v quyt nh globulin min dch thuc lp no. Chui nng c khong 440 acid amino v cng c hai vng. Vng c nh nm u carboxyl ( COOH), k hiu l C, c trt t acid amino ging nhau tt c globulin min dch thuc cng mt lp. Vng bin i nm pha u amin ( NH2), k hiu l V, trong trt t acid amin c mt s on hay thay i xen gia nhng on tng i n nh. Hai vng bin i ca chui nh v chui nng nm k nhau tham gia vo s hnh thnh v tr kt hp khng nguyn hay paratop ca phn t khng th mang tnh c hiu cho tng quyt nh khng nguyn. 5.3.1.3. Vng gp khc (domain), vng bn l v cc mnh phn t globulin min dch Cc cu ni disulfua va ni cc chui polypeptid vi nhau va ni cc acid amin nm xa nhau trong cng chui to nn nhng khc xon hoc cun hnh cu c gi l domain. Chui nh c 2 domain, chui nng c 4 domain. Edelman (1970) cho rng chc nng vng gp khc VH v VL l
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hp tc vi nhau to b mt ca v tr gn khng nguyn, cc vng khc lm nhim v trung gian cho cc chc nng ca khng th nh gn vi b th, gn vi th th cho Fc trn i thc bo, bch cu trung tnh,... Trong chui nng, vng gip gianh gia domain CH1 v CH2 c gi l vng bn l. Vng bn l c c tnh mm mi gip cho 2 cnh ca phn t khng th di ng c, c th m ra, khp vo t 0 180O, nh m khng th d dng kt hp vi khng nguyn.
Vng bn l cn l ni d b tc ng ca cc enzym tiu protein. Nu dng papain v pepsin ct phn t globulin min dch IgG, ta s thu c cc mnh khc nhau: Vi papain, thu 3 mnh: + 2 mnh Fab (Fragment of antigen binding), mi mnh gm mt chui nh v na chui nng c u amin. y l ni gn vi khng nguyn. + 1 mnh Fc (Fragment crystalizable), mnh ny c tnh c hiu khng nguyn, c kh nng gn ln b mt t bo, hot ho b th. Vi pepsin, thu 2 mnh: + Mnh ln Fab, c cu to gn nh mt phn t hon ton vi 2 ho tr do vn c kh nng phn ng vi khng nguyn. + Mnh nh Fc.
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Hnh 7.13. Cc mnh phn t Ig di tc dng ca papain v pepsin
5.3.2. Cc lp khng th C 5 lp globulin min dch: IgG, IgM, IgA, IgD, IgE. 5.3.2.1. IgG c chui nng , k hiu 22 hay 22 Chim 80% tng s Ig huyt thanh, vai tr chnh bo v c th, xut hin sau v thay th IgM trong chng nhim trng. L khng th duy nht truyn t m sang con qua nhau thai. Cn c vo s khc bit tnh khng nguyn vng c nh chui nng , IgG chia 4 phn lp: IgG1, IgG2, IgG3, IgG4. Chc nng ca IgG: + Opsonin ho: phn Fc ca IgG gn vi th th thch hp trn b mt i thc bo, phn Fab gn vi quyt nh khng nguyn, to iu kin cho i thc bo bt gi v tiu dit khng nguyn.
Hnh 7.14. Hin tng opsonin
Hot ho b th: IgG1, IgG2, IgG3 c kh nng hot ho b th theo con ng c in. Gy c t bo ph thuc khng th (Antibody Dependent Cellular Cytoxicity ADCC): phn Fab ca IgG gn vi t bo ch, phn Fc gn vi th th t bo K(Killer cell), to iu kin cho t bo gii phng cht c dit t bo ch (t bo mang khng nguyn l Tumor
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cell).
Hnh 7.15. Hin tng ADCC 1. T bo dit t nhin; 2. Khng th IgG; 3. T bo ch
Trung ho ngoi c t. Gy ngng kt t bo vi khun, trung ho virus: IgG gn trn b mt t bo lm cho vi khun kt thnh m, lm cho virus khng bm c vo th th t bo ch xm nhp.
5.3.2.2. IgM c chui nng , k hiu 22 hay 22 V cu trc: IgM do 5 n v c bn hp thnh (pentame), cc n v ni vi nhau bi chui peptid J, to ra 10 v tr kt hp khng nguyn. IgM chim 5 10% tng s Ig huyt thanh, l khng th xut hin sm nht trong nhim trng. IgM c 2 phn lp l: IgM1 v IgM2. Chc nng ca IgM: + Hot ho b th + Ngng kt vi khun.
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5.3.2.3. IgA c chui nng k hiu 22 hay 22 C 2 loi IgA: IgA trong huyt thanh v IgA tit. IgA huyt thanh chim 15 20% tng Ig huyt thanh ch yu dng monome, i sng ngn khong 6 ngy. Chc nng: Hot ho b th theo ng nhnh. IgA tit c trong sa, nc bt, nc mt, dch mi, dch tit ph qun, ng tiu ho, ng sinh dc, tit niu. IgA tit dng dime, hai phn t gn vi nhau phn Fc nh chui peptid J v mnh tit. Mnh tit l sn phm ca t bo biu m c tc dng bo v IgA trc enzym phn gii protein v gip vn chuyn IgA tit vo dch tit. Chc nng IgA tit: Chng VSV trn b mt nim mc, ngn chng xm nhp su vo cc c quan.
5.3.2.4. IgD c chui nng k hiu 22 hay 22 Chim 0,2 1 % tng s Ig huyt thanh. Chc nng cn bit rt t. 5.3.2.5. IgE c chui nng k hiu 22 hay 22 Nng trong huyt thanh thp, tng ln khi d ng, nhim k sinh trng. Chc nng: Gy qu mn tc th, do phn Fc ca IgE gn vi th th thch hp trn b mt bch cu a kim v t bo mast. Khi khng nguyn xm nhp s gn vi phn Fab, dn n cc t bo trn c hot ho gii phng cc hot cht nh histamin, serotonin, locotrien gy dn mch, tng thm thnh mch.
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6. B TH COMPLEMENT
6.1. M u
B th l mt nhm protein huyt thanh thuc h min dch t nhin. Nm 1930, Bordet chng minh bng cc th nghim in vitro l huyt thanh ti ca d c gy min dch chng phy khun t c tc dng lm ngng kt vi khun ny sau lm tan ra. Tc dng ny s gim v mt khi un huyt thanh 56OC trong 30 pht. Nh vy, trong huyt thanh con vt c gy min dch c hai yu t tham gia vo tc dng lm tan vi khun: mt yu t chu c nhit gy ngng kt c hiu vi vi khun nhng cha lm cht vi khun, l khng th; yu t th hai c sn trong huyt thanh, b bt hot 56OC sau 30 pht, c tc dng lm tan vi khun sau khi b khng th ngng kt, ban u c gi l alexin sau gi l b th (complement) do tc dng b sung ca n. Gan l c quan sn xut mi thnh phn b th cho mu, tr C1 do biu m ng tiu ho v ng tit niu sn xut. Cc k hiu v quy c quc t: C l b th ton phn. C l b th c hot ho, c hot tnh enzym tc ng tip ln cc b th tip theo, v d: C3b B th b mt hot tnh c k hiu c ch i (inactivated), v d: iC3b. H thng b th gm 9 protein k hiu t C1 C9. C1 gm 3 thnh phn: C1q, C1r, C1s Nhiu thnh phn b th l tin enzym khi b phn ct thnh 2 mnh: mnh a v mnh b. Phc hp b th do nhiu mnh lin kt lit to thnh, v d: C4b2b3b.
6.2. Hot ho b th
Cc b th mun hot ng phi c hot ho. S hot ho theo kiu phn ng dy chuyn v theo trnh t nht nh. Mt b th khi c hot ho li c kh nng kch thch hot ho b th tip theo to ra nhng hot cht c tc dng sinh hc thc hin c ch bo v. 6.2.1. Hot ho theo con ng c in 6.2.1.1. Tc nhn hot ho L s kt hp c hiu khng th vi khng nguyn. Ch c khng th IgM, IgG1, IgG2, IgG3 c kh nng ny v y l nhng khng th c nng cao trong mu, v vy, hot ho b th theo con ng c in xy ra rt ph bin. 6.2.1.2. Cc bc hot ho S kt hp khng nguyn khng th. Hot ho C1. Hot ho C4 v C2 tch C4a v C2b to C4bC2a l enzym phn gii C3. Hot ho C3 tch C3a to C4bC2aC3b . 6.2.1.3. Kt qu To C4bC2aC3b (C5 convertase) l enzym phn gii C5. 6.2.2. Hot ho theo ng nhnh ng nhnh l mt ng hot ho khc, bn cnh ng c in, c th xy ra c lp hoc kt hp vi ng c in. 6.2.2.1. Tc nhn hot ho B mt t bo vi khun, t bo nhim virus, k sinh trng, t bo nhim nm.
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Mt s polysaccharid t nhin: zymosan, endotoxin,... Cc yu t tham gia gm: B, D v P (B l globulin; D l globulin, P l globulin). 6.2.2.2. Cc bc hot ho Bnh thng trong huyt thanh c mt nng thp C3b (do qu trnh hot ho thng trc C3 vi s tham gia ca yu t B v D). C3b bm vo b mt l, tc nhn hot ho xc tin mnh qu trnh kt hp vi yu t B to C3bBb , cht ny li tc ng ln C3 to BbC3b3b. 6.2.2.3. Kt qu To BbC3b3b (C5 convertase) l enzym tc ng ln C5.
6.2.3. S hnh thnh phc hp tn cng mng L qu trnh tip ni cho c hai con ng, bt u t s hot ho C5 do enzym C5convertase v kt thc C9 to phc hp tn cng mng (membrane attack complexMAC), phc hp ny chc thng mng t bo vi khun gy tan bo.
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Hnh 7.20. S tm tt s hnh thnh phc hp tn cng mng
6.3. Tc dng sinh hc ca hot ho b th

vch mt s vi khun c cc phn t mannose lin kt vi nhau, cc phn t ny c kh nng gn vi cht lectin ca c th di s h tr ca hai phn t protease huyt thanh. S kt hp ny cng dn n hot ho C4 v t qu trnh hot ho ging nh con ng c in.
7. MIN DCH QUA TRUNG GIAN T BO (Cell Mediated ImmunityCMI)

p ng min dch qua trung gian t bo l phng thc th hai, bn cnh p ng min dch dch th nhm loi tr khng nguyn l. Gi l p ng min dch qua trung gian t bo v mun gy min dch th ng th phi dng hn dch t bo m khng th dng huyt thanh (dch th).
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Tm quan trng ca p ng min dch t bo c th hin r rt trong cc trng hp: Cc bnh nhim trng do VSV k sinh trong t bo. Mn cm do tip xc. Cc phn ng ca c th chng ung th v loi tr mnh ghp. Mt s bnh t min.
7.1. Lympho T
L t bo ph trch p ng min dch t bo, c ngun gc t tu xng di chuyn n tuyn c chn v bit ho.
7.2. Chc nng ca lympho T

Nhn bit khng nguyn do lympho T c CD4 v CD8 ph trch. iu ho, kim sot mc p ng min dch do TH v Ts ph trch. Loi tr khng nguyn do Tc, TDTH v cc t bo dit ph trch. Ghi nh min dch. 7.2.1. Chc nng nhn bit khng nguyn Lympho T nhn bit khng nguyn cho ton h min dch, cn lympho B ch nhn bit khng nguyn cho p ng min dch dch th. 7.2.1.1. Vai tr ca CD4 v CD8 a) Vi khng nguyn ngoi bo Khng nguyn ngoi lai xm nhp vo c th c i thc bo bt gi, x l v trnh din ln b mt nh phn t MHC lp II. Phn t CD4 ca lympho bo T c th gn kt c hiu vi phn t MHC lp II, gip lympho T tip cn vi khng nguyn. b) Vi khng nguyn ni bo Khng nguyn l (protein ca virus hay ca t bo ung th) c trnh din ln trn b mt t bo ch (t bo mang khng nguyn ni bo) nh phn t MHC lp I. Phn t CD8 ca lympho bo T c th gn kt c hiu vi phn t MHC lp I, gip lympho T tip cn vi khng nguyn.
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7.2.1.2. Vai tr ca th th TCR (Tcell receptor) Phn t CD4 v CD8 gip lympho T tip cn ng t bo trnh din khng nguyn bng MHC lp II hay MHC lp I. Cn vic trc tip nhn din khng nguyn l do th th TCR, nh vic gn trc tip vi khng nguyn. C rt nhiu qun th lympho T mang loi th th TCR gip nhn ra nhng khng nguyn ph hp.
TCR l mt protein mng ca cc t bo lympho T, v cu trc gm 2 chui polypeptid v gn vi nhau bng nhng cu ni disulfua. TCR cng c 2 vng: Vng bin i (V) nm u amin ca mi chui v to nn v tr kt hp khng nguyn, vng ny c trt t acid amin bin i mnh, to nn s a dng ca th th TCR, v mi qun th lympho T ch nhn ra mt khng nguyn ph hp.
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Vng hng nh (C): Vng ny c trt t acid amin khng i, ging nhau tt c cc TCR. 7.2.1.3. Vai tr ca cc phn t kt dnh Cc cp lin kt phn t MHC lp II CD4, MHC lp I CD8, TCRKN, l nhng cp kt dnh quan trng. Ngoi ra cn rt nhiu cp kt dnh khc nh: trn b mt t bo trnh khng nguyn c phn t ICAM (Intercellular Adhere Molecule: phn t kt dnh lin t bo), trn phn t nhn bit khng nguyn c cc phn t LFA1 (Lympho Function Antigen: khng nguyn chc nng ca lympho bo) v cc CD (Class phn lp, diferenciation bit ho, y l cc protein b mt t bo lympho biu th s bit ho ca cc phn lp) lm nhim v kt dnh. Vai tr ca cc phn t kt dnh: Lin kt cht t bo trnh khng nguyn v t bo nhn bit khng nguyn, gip cho s nhn bit khng nguyn hiu qu hn, gip hot ho v kch thch lympho T tit cc hot cht (lymphokin). 7.2.1.4. Vai tr ca cc cytokin Cytokin l cc hot cht do t bo min dch tit ra khi tip xc vi khng nguyn c hiu, c nh hng ti hot ng ca cc t bo khc (cyto: t bo, kinin: hot cht). Cc hot cht do bch cu tit ra gi l leukin, do cc t bo lympho tit gi l lymphokin. Cc hot cht quan trng trong tng tc gia cc t bo c quy nh tn gi quc t l interleukin IL. Hin tm c t IL1, IL2,..., IL10. Trong qu trnh nhn bit khng nguyn ca lympho T c CD4 v CD8, khng th thiu vai tr ca cc interleukin. Vi TH, cc cp kt dnh CD4MHC lp II, TCR khng nguyn l tn hiu th nht, cn hin tng i thc bo tit IL1 tc ng ln TH l tn hiu th hai cn v cho TH nhn din khng nguyn v hot ho. Vi TC, cc cp kt dnh CD8MHC lp I,TCR khng nguyn l tn hiu th nht, cn tn hiu th hai cn v tc ng ln TC l IL2, gip TC nhn din khng nguyn v hot ho. 7.2.2. Chc nng loi tr khng nguyn 7.2.2.1. Vai tr ca lympho TDTH TDTH l phn nhnh ca lympho T c CD4. TDTH c kh nng nhn din khng nguyn ngoi lai do MHC lp II ca i thc bo trnh din, y l tn hiu th nht, tn hiu th hai cn v l s kch thch ca IL2 gip TDTH hot ho v phn chia tng sinh vi mt cao vng khng nguyn xm nhp. Cc t bo TDTH hot ho c kh nng tit ra cc lymphokin thu ht i thc bo, bch cu trung tnh ti to vim nhm tiu dit khng nguyn. vim do TDTH hnh thnh mun sau 48 72 gi, do TDTH c gi l lympho gy qu mn mun. 7.2.2.2. Vai tr ca TC Sau khi c hot ho bi 2 tn hiu, tin lympho T C c bit ho thnh T C hot ng c kh nng dit cc t bo mang khng nguyn ni sinh, cc t bo ghp d gen bng vic tit cc lymphotoxin gy c t bo. c im ca c ch gy c trc tip ny l T C tip xc trc tip v gn cht vo t bo, tit cht c ph hu cc thnh phn bn trong t bo trc khi mng t bo b v ra.
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7.2.2.3. T bo NK (Natural killer: t bo dit t nhin) Trn b mt t bo NK c th th vi interferon v IL2, y l cht gy hot ho NK, gip n hot ho tit cht c ph hu t bo ch. 7.2.2.4. T bo lympho B p ng min dch dch th do t bo lympho B ph trch i vi khng nguyn ph thuc tuyn c (a s khng nguyn trong cuc sng thuc loi ny), phi c s h tr ca TH. T bo TH sau khi nhn din khng nguyn s tit IL2 t hot ho, chc nng chi phi min dch dch th ca TH l vic tip tc tit IL4 l yu t sinh trng gip mt qun th t bo B tng sinh, tip TH tit IL6 h tr qun th t bo B ny bit ho, chn, tr thnh tng bo sn xut khng th c hiu. 7.2.3. Chc nng iu ho, kim sot min dch 7.2.3.1. Chc nng iu ho min dch ca TH TH chi phi cc hot ng ca cc t bo min dch nh vic tit IL2, IL4, IL6, gip cho s sinh sn v hot ho cc t bo hiu ng mc loi tr khng nguyn. 7.2.3.2. Chc nng kim sot min dch ca TS TS l phn nhm ca lympho T c CD8, n c vai tr c ch phn ng loi tr khng nguyn do TH pht ng nu n qu mnh. 7.2.4. Ghi nh min dch
7.3. Qu trnh hnh thnh CMI

Qu trnh hnh thnh min dch qua trung gian t bo bao gi cng c s tng tc gia qun th t bo lympho v i thc bo. 7.3.1. i thc bo trnh din khng nguyn cho lympho T Khng nguyn xm nhp vo c th (khng nguyn ph thuc tuyn c) c i thc bo n, x l v trnh din nhm quyt nh khng nguyn ra b mt nh MHC lp II. 7.3.2. Mn cm gy hot ho lympho T 7.3.2.1. Mn cm ln u (vim khng c hiu) i thc bo theo ng bch huyt ti hch gn nht, cc lympho T vng v tng sinh mnh m lm hch sng to ln. Trn b mt cc t bo T hnh thnh nhng receptor c hiu vi i thc bo nhn din khng nguyn. Sau khong 6 ngy, nhng t bo T ny c mn cm s ri hch ban u i ti cc hch khc, ti lch gy mn cm cho nhng t bo T khc, ng thi, to ra dng t bo T nh cho p ng min dch th pht khi khng nguyn xm nhp ln sau. Nh vy, mt qun th t bo T trong hch, lch, tun hon c mn cm sn sng nhn din khng nguyn l v tiu dit
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chng. 7.3.2.2. Mn cm ln sau (vim c hiu) Nu khng nguyn gy mn cm ln u vo li ln sau, th ch cn mt lng nh, sau khong 6 8 gi phn ng vim hnh thnh v t ti a sau khong 48 72 gi.
7.4. Mt s hin tng CMI

7.4.1. Hin tng Koch Robert Koch l nh bc hc c, l ngi u tin tim vi khun lao vo di da chut lang bnh thng. Sau 10 14 ngy, ti ch tim xut hin mt cc rn, sau v ra to mt lot khng lnh c v chut cht. Nhng nu tim vi khun lao cho chut lang b nhim lao trc , th sau 2 ngy xut hin vim cng c thm nhim i thc bo v lympho bo, vim bong da to lot nng v chng lnh, chut khng cht. Th nghim ca Koch cho thy chut lang nhim lao c p ng min dch chng vi khun. p ng ny gip c th chut khu tr vi khun, khng cho lan to v tiu dit chng.
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7.4.2. Hin tng bong mnh ghp Ly mnh da ca chut A ghp cho chut B (cng loi nhng khng thun chng) trong iu kin v trng. Trong vi ngy u mnh da vn hng ho bnh thng, t ngy th 3 n th 9 th c biu hin mnh da tm i, hoi t xut hin v lan rng, ri da kh v bong ra, l ch ghp b vim ty. l hin tng bong mnh ghp. Tip li ly da chut A ghp cho chut B th mnh ghp ln sau ngy cng bong nhanh hn.
7.5. Cc bin php nh gi CMI

Test trong da: a mt lng nh v thch hp khng nguyn c hiu vo trong da, nu c th c p ng min dch t bo th ti ni tim din ra s tng tc gia khng nguyn v cc lympho T mn cm dn n s hnh thnh cc lymphokin thu ht, hot ho i thc bo, to mt phn ng vim sau 48 72 gi, gi l qu mn mun. Mt s test trong da thuc loi qu mn mun pht hin min dch t bo nh: test Mantoux dng khng nguyn tuberculin, nh gi min dch chng lao. Test Mitsuda dng khng nguyn lepromin nh gi min dch chng vi khun phong,... Xc nh s lng, t l lympho T trong mu.
8. MIN DCH CHNG NHIM VI SINH VT

Trong cuc sng, con ngi lun phi tip xc vi nhiu cn nguyn gy bnh l cc VSV. t bo v, c th s dng cc hnh thc p ng min dch t nhin v min dch c hiu. tn ti v pht trin, cc VSV cng s dng nhiu hnh thc n trnh min dch.
8.1. Min dch chng vi khun

8.1.1. Min dch chng vi khun ngoi bo Vi khun ngoi bo sng v nhn ln bn ngoi t bo ch, gy bnh bng c t v to ra phn ng vim dn n hu hoi t chc. p ng min dch ca c th l trung ho c t, loi tr vi khun. 8.1.1.1. C ch bo v t nhin chng vi khun ngoi bo a) Thc bo Hin tng thc bo c thc hin bi bch cu trung tnh, i thc bo t chc, monocyte. Kh nng thc bo b gim st khi c lc ca vi khun mnh. b) Hot ho b th Ni c t ca vi khun Gram() l mt tc nhn gy hot ho b th theo con ng nhnh, kt qu to c C3b gy opsonin ho gip tng cng thc bo, to phc hp tn cng mng gy tan bo, cc sn phm hot ho b th nh C3a, C5a lm tng cng phn ng vim nhm loi tr vi khun. c) Gii phng cytokin Ni c t ca vi khun Gram() c kh nng kch thch i thc bo, cc t bo ni mc mch sn xut cytokin v cc cht gy vim hot ho cc t bo min dch, gip bch cu bm dnh, xuyn mch lm nhim v thc bo. 8.1.1.2. C ch bo v c hiu Min dch th l p ng bo v chnh chng vi khun ngoi bo: + Khng nguyn khng ph thuc tuyn c trc tip kch thch lympho B hot ho sn xut khng th c hiu. + Khng nguyn ph thuc tuyn c c i thc bo bt gi, x l, v trnh din cng phn t MHC II cho t bo TCD4+, cc t bo ny s hot ho v tit lymphokin, gip lympho B sn xut
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khng th c hiu. Vai tr ca khng th: + Trung ho c t lm mt tnh c, ngn khng cho c t bm vo t bo. + IgA tit ngn cn vi khun bm vo nim mc ngn khng cho chng xm nhp c th. + Tng cng thc bo nh opsonin ho: cc khng th gn c hiu vi khng nguyn ti phn Fab, cn phn Fc gn th th ca C3b hoc vi th th trn b mt i thc bo. Nh opsonin ho m vi khun nhanh chng b tiu dit v o thi. 8.1.1.3. S n trnh p ng min dch ca vi khun ngoi bo Thay i cc protein b mt c kh nng bm chc vo b mt t bo tip cn v xm nhp su. Cc vi khun to v bc chng thc bo, c ch hot ho b th. Ngoi c t gy c cc t bo thc bo. Bin i khng nguyn b mt to khng nguyn mi lm mt hiu lc min dch c. 8.1.2. Min dch chng vi khun ni bo Vi khun ni bo sng v nhn ln trong t bo ch, v d nh trc khun lao, phong,... Cc khng th dch th lu thng khng tip cn c, do cc c ch p ng min dch chng vi khun ni bo khc vi c ch chng vi khun ngoi bo. 8.1.2.1. C ch bo v t nhin Thc bo l hnh thc bo v t nhin ch yu, nhng nhiu trng hp vi khun khi b t bo thc bo nut nhng khng b tiu dit m vn sng v tng sinh ngay trong cc t bo ny. 8.1.2.2. C ch bo v c hiu p ng min dch qua trung gian t bo l hnh thc bo v chnh. i thc bo bt, nut vi khun khng tiu dit c chng nhng vn trnh din c quyt nh khng nguyn ca vi khun cho cc t bo lympho TCD4. Cc t bo lympho ny s tng cng sn xut lymphokin kch thch i thc bo tiu dit vi khun. Nhim khun ni bo l vn phc tp, ngoi tnh khng nguyn, ng vo ca vi khun, cn c kh nng khng, tnh phn ng ca c th l vn cn c nghin cu thm. 8.1.2.3. S n trnh p ng min dch Vi khun ni bo ln trnh p ng min dch ca c th bng bin php chng li hin tng thc bo, v d, trc khun lao sau khi b i thc bo bt, nut vo phagosom th chng phn ng bng cch: c ch ho nhp ca phagosom v lysosom. Khng li cc enzym tiu trong phagolysosom. c thng mng phagosom thot vo bo tng trc khi c s ho nhp ca phagosom v lysosom.
8.2. Min dch chng virus

Virus l loi VSV sng ni bo, chng s dng b my ca t bo sn xut ra cc protein l gy p ng min dch dn n tn thng t bo, hoc gy chuyn bin t bo ch thnh t bo c tnh. Trc s tn cng nh vy, c th t bo v bng cc c ch sau: 8.2.1. C ch bo v khng c hiu Tng sn xut interferon t t bo nhim, cht ny c ch s nhn ln ca virus v hn ch s xm nhp ca virus vo cc t bo ln cn. T bo NK tng hot ng dit t bo nhim virus. Ngoi ra cc t bo thc bo, b th cng tham gia vo hot ng nhm tiu dit virus.
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8.2.2. C ch bo v c hiu Min dch c hiu chng virus bao gm c min dch dch th v min dch t bo. Khng th dch th c hiu c vai tr quan trong trong giai on sm khi virus cha xm nhp vo t bo. Cc IgA tit ngn cn cc virus tn cng theo ng nim mc, cc IgM v sau l IgG ngn cn virus bm dnh vo b mt t bo ch. Khi c khng th dch th, t bo NK c tng cng hot ng ph thuc khng th nhm dit t bo nhim virus (hin tng ADCC). Khng th dch th c vai tr quan trng trong giai on ban u, nhng khng th loi tr c virus khi n xm nhp vo t bo. Vy, c ch chnh ca min dch c hiu chng virus l min dch t bo vi vai tr ca T c, tc TC. Cc t bo T c mang CD8+ nhn bit khng nguyn virus trong s kt hp vi phn t MHC I. T c c hot ho s kch thch cc enzym, cc cytokin hot ng hn ch s xm nhp, ly gii t bo nhim, tiu dit virus.
Min dch c hiu chng virus l vn rt phc tp hin nay vn ang c tip tc nghin cu. Bn cnh tc dng bo v tch cc, trong mt s trng hp, p ng min dch li gy tn thng c th do to thnh phc hp min dch lng ng trn b mt m, hoc do ly gii mnh cc t bo nhim dn n hoi t m... 8.2.3. S n trnh p ng min dch ca virus Thay i khng nguyn b mt nh virus cm, nh vy m chng n trnh c khng th c hiu hnh thnh, do lm mt tc dng ca khng th. Lm tn thng cc t bo c thm quyn min dch nh HIV tn cng t bo TCD4+, l t bo quan trong ca h min dch vi chc nng nhn bit khng nguyn l, hot ho cc t bo min dch khc.
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9. VACCIN V HUYT THANH MIN DCH
9.1. Vaccin
9.1.1. Nguyn l Dng vaccin l a vo c th khng nguyn c ngun gc t VSV gy bnh hoc VSV c cu trc khng nguyn ging VSV gy bnh c bo ch m bo an ton cn thit, lm cho c th t to ra tnh trng min dch chng li tc nhn gy bnh. C th c c min dch sau khi dng vaccin l kt qu ca s p ng min dch i vi cc thnh phn khng nguyn c trong vaccin. Tu theo tng loi vaccin m c th c min dch th dch hay min dch qua trung gian t bo hoc phi hp c hai loi. Ch c nhng bnh truyn nhim sau khi ngi mc bnh khi, c th thu c min dch bo v mi c kh nng sn xut vaccin. 9.1.2. Phn loi vaccin 9.1.2.1. Theo ngun gc Vaccin VSV cht: Nui cy VSV gy bnh c c lc mnh trong mi trng thch hp ly khun lc. Dng cc nhn t l hc hoc ho hc git cht VSV nhng vn cn tnh khng nguyn, v d: vaccin phng bnh t. Vaccin VSV sng l nhng VSV c lm mt c lc nhng vn cn tnh khng nguyn. C th nui cy VSV trong nhng iu kin nht nh, hoc cy chuyn nhiu ln mi trng, nh nui cy vi khun lao trong mi trng mt b. Cng c th tim truyn qua ng vt nhiu ln nh chng virus u ma qua b c vaccin phng bnh u ma. Cc vaccin VSV sng phi c m bo thun khit v mt di truyn, ngha l nhng VSV khng bao gi c th tr li dng gy bnh ban u. V d: vaccin BCG phng lao, vaccin Sabin phng bi lit. 9.1.2.2. Theo thnh phn vi sinh vt Vaccin gii c t l vaccin c sn xut t ngoi c t ca VSV c lm mt c lc bng cc nhn t l hc, ho hc nhng vn gi c tnh khng nguyn. V d: vaccin gii c t bch hu, un vn. Vaccin v polysaccharid: V d: vaccin ph cu, haemophilus influenza typ b. Vaccin khng nguyn b mt: Vaccin vim gan B sn xut t huyt tng. Vaccin ADN ti t hp: Vaccin vim gan B sn xut t nm men. 9.1.2.3. Theo hiu lc min dch Vaccin n gi: Vaccin c sn xut t mt chng VSV, do ch c tc dng phng nga mt bnh. V d: vaccin phng bnh lao, bi lit. Vaccin a gi: vaccin gm nhiu loi khng nguyn cng mt lc c a vo c th phng nhiu bnh vi iu kin cc khng nguyn ny khng c ch ln nhau. V d: vaccin bch hu, un vn, ho g. 9.1.3. Nguyn tc s dng vaccin 9.1.3.1. Phm vi tim chng rng, t t l cao Phm vi tim chng c quy nh tu theo tnh hnh dch t ca tng bnh. Ni chung dng vaccin cng rng ri cng tt nhng cn ch n kh nng kinh ph, ch nhng vng ng dn c, vng trng im thng c dch xy ra.
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T l tim chng: Nhng khu vc c bnh truyn nhim lu hnh th tim chng phi t trn 80% i tng cm th mi c kh nng ngn nga c dch. Tim chng 50 80% th nguy c dch vn xy ra, di 50% th khng ngn nga c dch. 9.1.3.2. i tng dng vaccin Nhng ngi c iu kin tip xc vi VSV gy bnh m cha c min dch u c dng vaccin. Ring tr em sau khi ht min dch th ng do m truyn cho th nguy c mc bnh rt cao, cn c tim chng mt cch trit bi v tr em cng nh, bnh nhim VSV cng nng v t l t vong cng cao. i vi ngi ln i tng tim chng t hn, thng ch tim chng cho nhng ngi c nguy c cao. 9.1.3.3. iu kin sc kho m bo vaccin c iu kin gy c min dch, ni chung nn dng cho nhng ngi kho mnh. Vi mi loi vaccin c din chng ch nh ring song khng c dng vaccin cho cc i tng sau: Nhng ngi ang st cao, tuy nhin mt s trng hp st nh vn dng c vaccin. Nhng ngi ang b bnh d ng, ngi c c a d ng hoc gia nh c tin s d ng khi dng vaccin cn c theo di cn thn. + Vaccin VSV sng gim c lc khng c dng cho nhng ngi suy gim min dch, ngi ang dng thuc gy suy gim min dch, nhng ngi mc bnh c tnh hoc ph n c thai. 9.1.3.4. Thi gian dng vaccin Min dch do vaccin i hi phi c thi gian nht nh, thng th phi sau 7 10 ngy mi gy c min dch. V vy, mun phng c dch phi dng vaccin trc ma dch thng xy ra. Hiu gi khng th t cao nht sau khong 2 tun, l p ng min dch tin pht. Khong cch gia cc ln dng vaccin: Tu theo tng loi vaccin m c th dng mt ln hay nhiu ln. i vi nhng vaccin phi dng nhiu ln th khong cch tt nht gia cc ln l 1 thng. Nu khong cch ny ngn hn th mc d dng ln sau nhng kt qu p ng min dch vn ch nh tin pht. Nhng nu dng ln 2 sau ln 1 hn 1 thng th hiu qu min dch vn c m bo. Tuy nhin, khng nn ko di thi gian gia cc ln tim chng v c th b mc bnh trc khi vaccin c dng y . Thi gian dng nhc li: Khi dng nhc li vaccin, thi gian c min dch s ngn li, hiu gi khng th s t cao nht ch sau mt s ngy nh nhng t bo lympho c tr nh min dch. l kt qu ca p ng min dch th pht. Thi gian dng nhc li tu theo tng loi vaccin, ph thuc vo thi gian duy tr c tnh trng min dch c hiu lc bo v ca mi loi vaccin, v d: u ma 5 nm, bi lit 3 nm, thng hn 1 nm, t 6 thng. Sau thi gian tn ti min dch, cn c dng nhc li vaccin 1 ln. Vi ln ny, c th s p ng min dch nhanh hn v mnh hn mc d khng th ca ln trc ch cn rt t. 9.1.3.5. Liu lng: Tu theo tng loi vaccin v ng a vaccin vo c th m dng liu lng thch hp. Liu lng qu thp s khng kh nng kch thch c th p ng min dch. Ngc li, liu lng qu ln s dn n tnh trng t lit min dch c hiu i vi nhng ln tim chng tip theo (h thng min dch bnh thng ca c th khng c phn ng chng li cc khng nguyn). 9.1.3.6. ng dng vaccin Tim di da: a s cc loi vaccin a vo c th bng ng ny, phng php ny c hiu qu chc chn hn nhng d gy phn ng hn, liu vaccin nhiu so vi tim trong da 0,5ml. Tim trong da: Phng php tim trong da ch cn mt lng vaccin nh (0,1ml), t gy phn ng nhng cn lu phi tim ng k thut, nu khng m bo k thut th tc dng gy min dch km. Tim bp: Hin nay, c mt s vaccin phi tim bp mi t b bin chng v hiu lc min dch cao nh vaccin tam lin bch hu, un vn, ho g, hoc vaccin gii c t un vn. ng chng: l hnh thc rch da a vaccin vo nh chng vaccin phng bnh u ma.
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y l phng php th s v c in nht. ng ung: l ng a vaccin vo c th d dng v tin li nht, khng gy phn ng. Tuy nhin ch thc hin c vi nhng vaccin khng b ng tiu ho ph hu. ng ung kch thch min dch tit ti ch mnh hn nhiu so vi ng tim. V d: vaccin Sabin phng bi lit. 9.1.3.7. Cc phn ng ph do dng vaccin Khi dng vaccin, mt s ngi c th xy ra phn ng ph nh: Ti ch: c th au, hi sng hoc ni cc , hin tng ny mt i sau mt vi ngy. Nu tim chng khng m bo v khun c th gp vim nhim c m, lot ch tim. Ton thn: Thng gp nhiu nht l st (10 20%), st thng ht sau vi ngy. C th gp t l rt thp b co git, sc phn v. Tuy nhin, mc nguy him do vaccin gy ra nh hn rt nhiu so vi mc nguy him do bnh nhim trng gy ra. V vy, rt cn thit phi dng vaccin phng bnh. Nu sau khi dng vaccin 2 3 ngy m cc phn ng vn cn, phi n y t kim tra. 9.1.3.8. Bo qun vaccin Vaccin l mt sinh phm nn d b hng nu khng c bo qun ng. Mi loi vaccin c yu cu bo qun ring nhng ni chung phi bo qun iu kin kh, ti v lnh. Nhit cao, nh sng v ng lnh ph hu nhiu vaccin. Nhit bo qun tt t 2 8OC. Mi loi vaccin c thi hn s dng nht nh c ghi trn nhn nn cn c kim tra trc khi s dng. 9.1.4. Lch tim chng Cn c vo c im dch t hc ca cc bnh truyn nhim, tui tt nht cho vic tim chng, thi gian nguy c mc bnh, s p ng min dch ca cc i tng c tim chng, kh nng cung cp vaccin lm c s xy dng lch tim chng (bng 7.1).
9.2. Huyt thanh min dch
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9.2.1. Nguyn l Dng huyt thanh min dch l a vo c th mt loi khng th c ngun gc t ngi hay ng vt, lm cho c th c ngay khng th c hiu chng li tc nhn gy bnh. y l min dch th ng nn chng ht, ch tn ti trong c th vi ngy. 9.2.2. Nguyn tc s dng 9.2.2.1. i tng Huyt thanh c dng iu tr v phng bnh cho nhng bnh nhn nhim VSV hay c t cp tnh. Cn a ngay khng th trung ho tc nhn gy bnh. Huyt thanh ch c hiu lc vi nhng bnh m c ch bo v ch yu nh min dch dch th. V d: huyt thanh chng un vn SAT, huyt thanh chng bch hu SAD, huyt thanh khng di. Khi dng huyt thanh thng phi hp vi khng sinh dit khun v vi vaccin gy min dch ch ng bo v lu di hn. 9.2.2.2. Liu lng Tu theo tng la tui v mc ca bnh m s dng liu khc nhau, trung bnh 0,1 1ml/kg cn nng. Mt s huyt thanh c tnh theo n v nh khng c t un vn, bch hu, trung bnh l 250 n v cho 1 ln. 9.2.2.3. ng a huyt thanh vo c th a s cc loi huyt thanh c tim bp. i vi nhng huyt thanh c ngun gc t ngi, c tinh ch t tiu chun cao c th tim tnh mch nhng cng rt hn ch. Khng c tim tnh mch nhng huyt thanh c ngun gc ng vt. 9.2.2.4. phng phn ng phng phn ng do huyt thanh, trc khi dng cn ch : Hi bnh nhn dng huyt thanh ln no cha; Thn trng khi dng t ln th 2 tr i v t l phn ng cao hn so vi ln th nht. Lm phn ng gii mn cm: Pha long huyt thanh 10 ln vi nc mui sinh l v khun. Tim 0,1ml vo trong da, 30 pht sau khng c hin tng qung th c th tim huyt thanh. Nu c qung ti ni tim th khng nn tim. Trong trng hp bt buc phi dng th chia nh tng liu tim dn, cch nhau 20 30 pht. Trong qu trnh truyn huyt thanh phi theo di lin tc v chun b y cc iu kin x l kp thi nu c phn ng xy ra. 9.2.3. Cc phn ng ph khi dng huyt thanh 9.2.3.1. Ti ch Ni tim c th au, mn , ht sau vi ngy v khng gy nguy him. 9.2.3.2. Ton thn Bnh nhn rt run, kh th, au khp, c th nhc u v nn. Nng nht l sc huyt thanh c th xut hin sau khi tim ln th nht 10 14 ngy v lc c th sinh khng th chng li, hoc xy ra ngay sau khi tim, hoc mt vi ngy sau khi tim huyt thanh ln th 2. Triu chng thng gp l kh th, nga v ni m ay ton thn, au bng, b i.
10. CC PHN NG MIN DCH TRONG CHN ON VI SINH VT
10.1. Mc ch
Cc phn ng min dch dng trong chn on VSV ch yu l cc phn ng kt hp gia khng nguyn v khng th nhm mc ch:
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10.1.1. Pht hin khng nguyn Dng khng th bit trc pht hin hoc chun khng nguyn VSV. K thut ny thng c dng trong chn on cc bnh nhim trng: Sau khi nui cy phn lp c VSV t bnh phm ca bnh nhn cho kt hp vi khng huyt thanh mu c khng th bit trc xc nh tn VSV. K thut ny c th dng khng th mu pht hin trc tip khng nguyn ca VSV c trong bnh phm m khng cn phi nui cy phn lp. Ngoi vic chn on bnh nhim trng, k thut pht hin khng nguyn cn c dng trong nh loi VSV, tm hiu v cu trc khng nguyn VSV khi cho kt hp vi khng huyt thanh mu. 10.1.2. Pht hin khng th hoc xc nh hiu gi khng th K thut pht hin khng th thng c dng trong chn on cc bnh nhim trng m c th c sinh ra khng th chng li VSV gy bnh. Dng cc khng nguyn mu bit trc cho kt hp vi huyt thanh bnh nhn pht hin khng th, v vy cn c gi l phn ng huyt thanh hc. Vic pht hin khng th cng thng c p dng trong nghin cu iu tra dch t hc ca cc bnh nhim trng thng qua vic iu tra khng th trong huyt thanh ca nhng ngi trong phm vi nghin cu. Ngoi ra, pht hin khng th cn dng nghin cu p ng ca c th i vi khng nguyn VSV, v d: Khi dng th nghim mt loi vaccin no , phi nh gi hiu lc p ng min dch bng cch xc nh hiu gi khng th. D l mc ch xc nh khng nguyn hay khng th th kt qu ca phn ng u c nh gi thng qua s hnh thnh phc hp khng nguyn, khng th. Phc hp ny c th quan st trc tip hoc nh gi gin tip thng qua mt s hin tng khc.
10.2. Bn cht ca s kt hp khng nguyn, khng th

S kt hp gia khng nguyn v khng th l s gn chnh xc gia v tr khng nguyn v v tr khng th. S kt hp ny mang tnh c hiu, ngha l khng nguyn ch kt hp vi khng th m n kch thch c th sinh ra, khng th ch kt hp vi khng nguyn kch thch c th sinh ra n. Tnh c hiu ny do cu hnh khng gian ca v tr khng nguyn v v tr khng th quyt nh, nh kho vi cha kho.
10.3. Cc phn ng kt hp khng nguyn, khng th

10.3.1. Cc phn ng to thnh ht Phn ng kt hp khng nguyn, khng th nh gi kt qu da vo s hnh thnh ht, nhng ht ny chnh l phc hp khng nguyn, khng th c hnh thnh khi khng nguyn v khng th c hiu c t l thch hp. C th quan st nhng ht ny bng mt thng hoc knh lp. 10.3.1.1. Phn ng kt ta Nguyn l: Phn ng kt ta l s kt hp gia khng nguyn ho tan (khng nguyn tm phn t) vi khng th tng ng, to thnh cc ht c th quan st bng mt thng. Cc loi phn ng kt ta: + Phn ng kt ta trong mi trng lng: Trn dung dch khng nguyn vi dung dch khng th theo mt t l nht nh trong ng nghim hoc trn phin knh. Phc hp khng nguyn khng th l nhng ht l lng hoc lng xung y ng nghim hay trn b mt hn dch. + Phn ng kt ta trong mi trng gel: Mi trng gel c dng l thch, c nhiu k thut kt ta trong gel thch, k thut khuch tn trong ng nghim, trn phin knh, trong a petri. Khng nguyn v khng th khuch tn ngc chiu nhau v kt hp c hiu to nn cc ng ta, hoc l khng th c ho u trong gel thch, khng nguyn c kh nng khuch tn trong mi trng thch, khi khng nguyn gp khng th tng ng s kt hp vi nhau v trong mi trng xut hin ng ta. Nu trong thch c
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ho nhiu loi khng th v dung dch khng nguyn c nhiu loi th trong thch s xut hin nhiu ng ta. 10.3.1.2. Phn ng ngng kt Nguyn l l s kt hp gia khng nguyn hu hnh (t bo hoc vt th c kch thc tng ng) vi khng th (ngng kt t) to thnh phc hp khng nguyn, khng th di dng nhng ht ngng kt c th quan st bng mt thng. iu kin hnh thnh mng li ngng kt: Ngoi tnh c hiu ca khng nguyn v khng th, c hin tng ngng kt phi c cc iu kin: + Khng nguyn v khng th phi a gi (c nhiu v tr kt hp). + Khng nguyn v khng th c nng tng ng nhau.
Cc loi phn ng ngng kt: + Phn ng ngng kt trc tip (ngng kt ch ng): Thnh phn khng nguyn ca chnh t bo vi khun, kt hp vi khng th c hiu to thnh mng li ngng kt. Trong thc t, phn ng ngng kt trn lam knh thng c s dng ph bin nht trong cc phng xt nghim VSV.
Hnh 7.28. Phn ng ngng kt trc tip gia vi khun v KT c hiu
+ Phn ng ngng kt gin tip (ngng kt th ng): Trong cc phn ng ngng kt gin tip, khng nguyn dng ho tan c gn ln nn mn (thng l hng cu hoc ht cht do nh ht latex). Khi gp khng th c hiu, khng nguyn s kt hp vi khng th, hin tng ngng kt xy ra do nn mn t li vi nhau mt cch th ng. Phn ng ngng kt gin tip tin hnh n gin, nhanh, nhy cao, c hiu cao nn c dng rng ri trong chn on vi sinh.
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10.3.2. Phn ng da vo hot ng sinh hc ca khng th C 2 loi phn ng thng c s dng trong chn on VSV l phn ng trung ho v phn ng kt hp b th. 10.3.2.1. Phn ng trung ho Nguyn l: Khi khng nguyn l vi khun, virus v khng th c hiu kt hp vi nhau th khng th c kh nng trung ho c lc virus v c t ca vi khun lm mt kh nng gy bnh ca chng. Cc loi phn ng trung ho: C nhiu phn ng trung ho c tin hnh trn ng nghim (invitro) hoc trn sc vt (invivo) chn on cc bnh do vi khun, virus, xc nh khng th trong huyt thanh bnh nhn hoc xc nh kh nng gy bnh ca vi khun, virus. Mt s phn ng thng dng l: + Phn ng trung ho virus: Phn ng ny thng dng chn on virus. C s ca phn ng l: Mt s virus c kh nng gy hu hoi t bo hoc git cht ng vt th nghim. Khi cho virus ny kt hp vi huyt thanh c hiu s lm mt cc kh nng trn. Nguyn l ca phn ng c tm tt s sau: Virus + t bo T bo b hoi t. Virus + khng th c hiu + t bo T bo khng b hoi t. + Phn ng trung ho trn sc vt: Nguyn l ca phn ng l khng th c hiu s trung ho khng nguyn l ngoi c t ca mt s loi vi khun c tim vo sc vt, lm cho sc vt khng b mc bnh. V d: xc nh vi khun bch hu hay gi bch hu, ngi ta tim vi khun ny vo hai con chut lang, mt con c tim khng c t bch hu, mt con khng. Nu con chut c tim khng c t bch hu vn sng, con chut kia cht th vi khun c xc nh l bch hu. 10.3.2.2. Phn ng gy ly gii t bo (kt hp b th) Nguyn l: Khng th c hiu vi s tham gia ca b th s gy ly gii t bo vi khun hoc mt s t bo ng vt. Phn ng kt hp b th thng c dng nhiu trong chn on.
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10.3.3. Phn ng dng khng nguyn hoc khng th nh du Cc phn ng thuc loi ny c nguyn l chung l khng nguyn hoc khng th c xc nh nh cht nh du c gn vi khng th hoc khng nguyn. Cht nh du khng c lm thay i hot tnh min dch ca khng nguyn v khng th. 10.3.3.1. Phn ng min dch hunh quang Cht nh du trong phn ng ny l cht mu hunh quang, c kt qu phn ng knh hin vi hunh quang. Phn ng min dch hunh quang trc tip: Phng php ny dng khng th gn hunh quang pht hin khng nguyn. Bng phng php min dch hunh quang trc tip c th pht hin nhanh khng nguyn VSV trong bnh phm v thng c dng trong chn on virus.
Phn ng min dch hunh quang gin tip: Phng php ny dng khng nguyn mu v khng khng th (KKT) gn hunh quang pht hin khng th.
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10.3.3.2. Phn ng min dch phng x (RIA Radioimmunoassay) Trong phn ng min dch phng x, cht nh du l cht ng v phng x, phc hp khng nguyn khng th c pht hin nh cht ng v phng x. Pht hin ni xy ra phn ng kt hp khng nguyn khng th nh hin tng pht x. 10.3.3.3. Phn ng min dch men ELISA (Enzym Linked Immunosorbent Assay) Dng khng th (KT) hay khng khng th (KKT) c gn enzym. Khng th kt hp vi khng nguyn to thnh phc hp khng nguyn khng th ng thi c mt ca enzym gn trn . Sau , cho thm mt c cht thch hp. Enzym tc ng ln c cht lm thay i mu mi trng. Cc men thng c s dng l peroxidase, oxydase, betaglactosidase. C nhiu k thut ELISA, sau y l mt s k thut thng dng: K thut dng khng khng th (KKT) gn enzym pht hin khng th
K thut dng khng khng th (KKT) gn enzym pht hin khng nguyn
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10.4. Nhn nh kt qu phn ng kt hp khng nguyn khng th

10.4.1. nh tnh nh tnh l k thut ch cho bit trong mu xt nghim c khng th hoc khng nguyn hay khng. Kt qu c nh gi cc mc v k hiu: Dng tnh: (+++; ++; +). m tnh: (). Khng r: (+/). Trong mt s trng hp, ch cn nh tnh c gi tr chn on, l cc trng hp m khng nguyn hoc khng th ngi bnh thng khng c nh bnh giang mai. Tuy nhin, c nhng bnh m khng nguyn hoc khng th tm thy c ngi bnh thng th phi nh lng mi c gi tr chn on, v d: Tm khng th trong bnh thp khp do lin cu nhm A. 10.4.2. nh lng Nng khng th trong huyt thanh cao hay thp c nh gi bng hiu gi khng th. Hiu gi khng th l pha long huyt thanh ln nht m phn ng cn dng tnh.
10.5. Chn on huyt thanh hc (Serolopy)

L phng php pht hin khng th trong mu bnh nhn chn on bnh nhim trng tng ng. nh gi c kt qu chn on, cn phi ly bnh phm 2 ln, ln th nht vo tun u ca bnh v ln th hai sau 7 10 ngy tm ng lc khng th. ng lc khng th l i lng c trng cho mc thay i hiu gi khng th theo thi gian. ng lc khng th l thng s gia hiu gi khng th ca mu huyt thanh ln th hai v ln th nht. ng lc khng th t nht phi bng 4 th mi c gi tr chn on bnh nhn ang mc bnh nhim trng. C th khng phi lm hai ln nu c quy nh hiu gi gii hn, t hiu gi gii hn tr ln c th coi l chn on dng tnh.
11. TNG QUT V QU TRNH IU HO MIN DCH

Cng nh mi qu trnh sinh hc, p ng min dch lun c iu ho m bo cn bng hot ng ca h min dch, gi hng nh ni mi. Nu s p ng cha lp tc c kch thch gia tng, nu p ng qu mc s b km hm gim bt v khi khng cn thit s ngng hot
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ng. S iu ho min dch bao gm c vic khng h min dch chng li cc khng nguyn ca bn thn nhm trnh mc cc bnh t min. Tm hiu qu trnh iu ho min dch c li ch quan trng trong vic iu tr cc bnh l min dch nh suy gim, qu mn, trong vic iu ch vaccin phng bnh c hiu lc. Tham gia kim sot v iu ho min dch l qu trnh rt phc tp vi cc yu t nh: khng nguyn, khng th, cc cytokin, mng li idiotyp khng idiotyp, s hp tc ca cc t bo lympho, h thn kinh ni tit. Di y chng ti ch gii thiu mt m hnh tng qut v hp tc t bo v iu ho min dch chng khng nguyn ph thuc tuyn c. Theo m hnh ny (hnh 7.35), cc khng nguyn ph thuc tuyn c khi xm nhp s b i thc bo bt gi, x l ri trnh din quyt nh khng nguyn trn b mt cho t bo T nh cc phn t MHC lp II. T bo lympho T cm ng (inducer T cellTI) khi u qu trnh iu ho bng vic tit yu t c tc dng hot ho i thc bo (macrophage activating factor MAF) lm i thc bo hot ho v tit IL1, cht ny tc ng tr li lympho TI, lc ny TI nhn 2 tn hiu kch thch: mt l quyt nh khng nguyn nm trong phc hp vi MHC lp II v hai l IL1. TI c hot ho s tng sinh v tit IL2, cht ny s tc ng ln cc t bo cng n nhn din quyt nh khng nguyn trn b mt i thc bo, l lympho T h tr, lympho T gy qu mn mun. Cc t bo ny sau khi nhn din khng nguyn s xut hin th th b mt tip nhn s kch thch ca IL 2, sau khi nhn 2 tn hiu chng c hot ho pht huy tc dng. Lympho T gy qu mn mun (TDTH ) c hot ho s tng sinh tit cc lymphokin tc ng ln cc bch cu ht (bch cu trung tnh,bch cu i kim, bch cu i toan) v i thc bo thu ht chng n ni c khng nguyn, hnh thnh phn ng vim kiu qu mn mun nhm loi tr khng nguyn. Lympho T h tr lympho B (TH) khi nhn 2 tn hiu kch thch s tng sinh v tit IL4, IL5, IL6 thc y lympho B bit ho thnh t bo plasma sn xut khng th. Tin lympho T c sau khi nhn din khng nguyn l trn b mt t bo ch th xut hin th th tip nhn tn hiu t IL2. Sau khi nhn 2 tn hiu, tin T c s bit ho thnh lympho T c (TC) c kh nng gy c trc tip t bo ch. Trong m hnh ny cho thy ng thi vi cc t bo TI, TH, TDTH, t bo TFR cng tng tc vi i thc bo nhn din khng nguyn v hnh thnh th th nhn kch thch t IL2. Sau khi nhn 2 tn hiu t bo ny s hot ho v kch thch cc lympho T c ch (TS). T c ch hot ng s tc ng tr li vi TI iu ho vic bi tit IL2 v tc ng ln cc loi TI, TH, TDTH lm gim hoc ngng p ng min dch. Tc dng c ch ca TS ch tc ng ln cc t bo trn giai on ang bit ho do vy khng lm p ng min dch dp tt ngay t u m ch khi p ng khi pht v xy ra mt mc no . M hnh trn y cha hon ton ng trong mi trng hp c th, song n cho php cho chng ta hnh dung cc bc xy ra trong qu trnh p ng min dch mt cch tng qut.
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CU HI LNG GI
1. Nu cc hnh p ng min dch t nhin ca c th. 2. Nu khi nim v c im c bn ca min dch c hiu. 3. Trnh by phn loi min dch c hiu. 4. Phn tch chc nng ca cc t bo tham gia vo qu trnh min dch. 5. Nu nh ngha v tnh cht ca khng nguyn. 6. Trnh by phn loi khng nguyn VSV. 7. Trnh by qu trnh tng sinh, bit ho t bo lympho B. 8. Nu nh ngha v hai c tnh quan trng ca khng th. 9. Trnh by cu trc, chc nng ca phn t khng th. 10. B th l g? Ti sao phi hot ho b th. Trnh by tc dng sinh hc ca hot ho b th. 11. Trnh by ngun gc v chc nng ca lympho T v qu trnh hnh thnh CMI. 12. Trnh by p ng min dch ca c th chng vi khun ngoi bo. 13. Trnh by p ng min dch ca c th chng vi khun ni bo. 14. Trnh by p ng min dch ca c th chng virus. 15. Nu nguyn l v nguyn tc s dng vaccin. 16. Trnh by phn loi vaccin. 17. Nu nguyn l v nguyn tc s dng huyt thanh. 18. Trnh by mc ch, nguyn l, cch nhn nh kt qu ca cc phn ng min dch.
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PHN TH BA
Chng 8
CC VI KHUN GY BNH THNG GP
1. CC CU KHUN GY BNH
Cu khun l nhng vi khun hnh cu, hiu kh tuyt i hoc hiu kh, k kh tu tin. Nhng vi khun gy bnh hiu kh, k kh tu tin c chia lm cc nhm sau da vo tnh cht bt mu: Cc cu khun Gram(+) thuc loi t cu (Staphylococcus), thuc loi lin cu (Streptococcus) v song cu (Diplococcus hay Streptococcus pneumoninae). Cc cu khun Gram() thuc loi Neiseria.
T CU
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(Staphylococci) T cu l mt trong nhng vi khun c pht hin sm. Nm 1878, Robert Koch m t t cu. Nm 1880, Louis Pasteur phn lp c t cu. Nm 1881, Ogston gy bnh thc nghim t cu. T cu phn b rng ri trong t nhin v thng k sinh trn da v cc hc t nhin ca ngi. Trong , nhiu loi l nhng t cu cng sinh khng gy bnh v mt s loi gy bnh gii hn. T cu thuc h Micrococcaceae, h ny c cc ging: Planococcus: khng gy bnh cho ngi. Micrococcus: gy bnh cho ngi. Staphylococcus: gy bnh cho ngi, ging ny c 13 loi, trong c 3 loi c vai tr quan trng trong y hc l: S. aureus (t cu vng), S. epidermidis (t cu da) gy nhim khun c hi v S. saprophyticus gy nhim khun tit niu.
1.1. T cu vng (Staphylococcus aureus)

1.1.1. c im sinh vt hc 1.1.1.1. Hnh th v tnh cht bt mu T cu l nhng vi khun hnh cu c ng knh 0,8 1m ng t li vi nhau thnh tng m nh chm nho, bt mu Gram(+). T cu thng khng c v, khng c lng, khng sinh nha bo.
Hnh 8.1. Staphylococcus aureus quan st di knh hin vi quang hc
1.1.1.2. Tnh cht nui cy T cu vng thuc loi d nui cy, pht trin d dng trn cc mi trng nui cy thng thng. Nhit thch hp l 10 45OC v nng mui cao ti 10%. Trong mi trng canh thang, vi khun pht trin mnh v lm c u mi trng, lu y c lng cn. Trong mi trng thch thng, sau 24 gi 37OC, to khun lc dng S, c mu vng chanh. Trong mi trng thch mu, t cu pht trin nhanh to khun lc dng S, tan mu hon ton. 1.1.1.3. Tnh cht ho sinh Cc enzym: T cu c h thng enzym phong ph, cc enzym dng trong chn on l: + Coagulase: C kh nng lm ng huyt tng ngi v ng vt khi c chng ng.
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y l tiu chun quan trng nht chn on v phn bit t cu vng. C 2 loi coagulase: mt loi tit ra mi trng gi l coagulase t do, mt loi bm vo vch t bo gi l coagulase c nh. Chng c tc dng to ra cc mu ng xung quanh t bo vi khun, do vy t cu vng trnh c tc dng ca khng th v hin tng thc bo. + Fibrinolysin (staphylokinase) l enzym c trng cho cc chng gy bnh ngi. Enzym ny lm tan cc mu v hnh thnh nhng vt tc mch nh to ra nhim khun di cn. + Hyaluronidase: Phn gii cc acid hyaluronic ca m lin kt gip vi khun lan trn vo m. + lactamase: Lm mt tc dng ca penicillin. + Catalase: Xc tc gy phn gii H2O2 O2 + H2O. + Desoxyribonuclease: Phn gii ADN. Ln men ng mannitol. 1.1.1.4. c t c t rut (Enterotoxin): a s t cu vng tit ra c t rut gy nhim c thc n v vim rut cp. Ngoi c t (Efoliatin toxin): Gy nn hi chng phng rp v trc l da tr em. c t gy hi chng shock nhim c (Toxic shock syndrome toxin). c t ny thng gp nhng ngi b nhim trng vt thng. Ngoi c t sinh m (Pyogenic exotoxin) c tc dng sinh m v phn bo lymphocyte. c t bch cu (Leucocidin) lm bch cu mt tnh di ng v b ph hu nhn. c t ny ch tc dng vi bch cu a nhn v i thc bo. N cng c tc dng lm hoi t da th. Dung huyt t (Hemolysin). C 4 loi dung huyt t: + Dung huyt t gy tan hng cu th, cu, gy hoi t da th v gy cht chut v th, gy hoi t t bo nui cy. + Dung huyt t gy tan hng cu ngi, cu, b. Liu cao gy cht th, hoi t t bo nui cy. + Dung huyt t gy tan hng cu ngi v nhiu ng vt. Gy hoi t nh da th v gy cht th. + Dung huyt t gy tan hng cu ngi, nga, th, cu, lm x cng da th, hoi t t bo nui cy. 1.1.1.5. Khng nguyn Acid teichoic: Acid ny gn vo polysaccharid vch t cu, y l mt thnh phn ca khng nguyn O l khng nguyn ngng kt v lm tng tc dng hot ho b th, cn l cht bm dnh ca t cu vo nim mc mi. Protein A l nhng protein bao quanh b mt vch t cu vng, 100% cc chng t cu vng c khng nguyn ny. Nhng chng t cu sn sinh nhiu protein A th tc dng thc bo gim i r rt. Polysaccharid: Mt s chng t cu c v th c khng nguyn ny. V c tc dng chng thc bo. Khng nguyn adherin (yu t bm): y l mt protein b mt ca t cu, c tc dng bm vo cc receptor c hiu ca t bo. 1.1.1.6. Sc khng T cu vng c sc khng vi nhit v ho cht cao hn cc loi vi khun khng sinh nha bo. B cht 80OC trong mt gi. T cu c kh nng gy bnh sau mt thi gian di sng ngoi mi trng. 1.1.1.7. Khng khng sinh Kh nng khng khng sinh ca t cu vng l mt c im rt ng quan tm. a s cc chng khng li penicillin G do sn sinh c penicillinase nh gen trn Rplasmid. Mt s chng khng li methicillin gi l methicillin resistant S. aureus (MRSA), do to ra nhng protein gn vo v tr
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tc ng ca khng sinh. Hin nay, mt s chng khng c vi cephalosporin cc th h, khng sinh c dng trong trng hp ny l vancomycin. 1.1.2. Kh nng gy bnh cho ngi 1.1.2.1. Nhim khun ngoi da T cu k sinh da v nim mc nn chng c th xm nhp qua cc vt thng hoc l chn lng gy nhim khun sinh m: mn nht, u inh, cc apxe, eczema, hu bi,... Mc nhim khun ph thuc vo s khng ca c th v c lc ca vi khun. Nhim t cu ngoi da thng gp tr em v ngi suy gim min dch. 1.1.2.2. Nhim khun huyt T cu vng l vi khun hay gy nhim khun huyt nht. Nhim khun thng xy sau nhng nhim khun tin pht, c bit l cc nhim khun ngoi da, t vi khun xm nhp vo mu. y l mt nhim trng nng, t mu, t cu n cc c quan khc gy cc apxe ( gan, phi, no, xng) c th gy vim tc tnh mch, t l t vong cao. 1.1.2.3. Nhim c thc n v vim rut cp Nhim c thc n thng do n ung phi c t rut hoc do t cu vng vn c tr ng rut, khi dng khng sinh ko di, vi khun ch bnh thng b tiu dit th t cu vng (khng khng sinh) c iu kin thun li pht trin nhanh v s lng tit c t rut gy bnh. Triu chng ng c rt cp tnh. Sau khi n phi thc n nhim c t vi gi, bnh nhn nn v tiu chy d di, phn nhiu nc. C th dn n sc do mt nc v in gii. 1.1.2.4. Vim phi Vim phi do t cu vng t gp. Thng xy ra sau vim ng h hp do virus hoc sau nhim khun huyt. C th tin pht do t cu vng, gp tr em, ngi gi v nhng ngi suy yu. T l t vong kh cao. 1.1.2.5. Nhim khun bnh vin Thng rt hay gp, nht l gy nhim trng vt m, vt bng,... t dn n nhim khun huyt, cc chng t cu bnh vin khng khng sinh rt mnh. Ngoi cc bnh thng gp trn, t cu cn c th gy hi chng phng rp da, vim da hoi t, sc nhim c do ph n s dng bng gc khng sch khi kinh nguyt. 1.1.3. Min dch Min dch thu c vi t cu t c vai tr bo v. p ng min dch qua trung gian t bo c th xy ra, nhng khng lm tng s dit khun. Min dch dch th cng xut hin chng li c t v enzym, nhng t cu t tip xc vi khng th do thng trong cc apxe, cc mu ng. 1.1.4. Chn on vi khun hc Bnh phm: Tu theo th bnh m ly bnh phm thch hp. Bnh phm phi ly ng v tr, ng thi gian v m bo v khun. Nhum soi trc tip: Phng php nhum soi cho php chn on s b khi nhn nh hnh th m khng c gi tr chn on quyt nh v trn da v nhiu b phn bnh thng cng c t cu k sinh nhng khng gy bnh. Nui cy v xc nh tnh cht sinh ho hc, y l phng php chn on xc nh t cu gy bnh: + Bnh phm l m, dch: Cy bnh phm vo mi trng thch mu. Sau 24 gi, nhn xt hnh thi khun lc, xem tnh cht tan mu. Nu nghi ng l t cu th cy chuyn sang cc mi trng xc nh kim tra cc tnh cht sinh ho hc ca t cu. + Bnh phm l mu: Ly 5 10ml mu tnh mch bng th thut v khun cy vo bnh mi trng c khong 100 150ml canh thang. t m 37OC, theo di hng ngy. Nu mi trng c th nhum soi, nu c t cu Gram(+) th cy chuyn sang mi trng thch mu v cc mi trng
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kim tra cc tnh cht sinh ho hc ca t cu. + Bnh phm l phn: cy vo mi trng Schapman, t m 37OC. Sau 24 48 gi, chn khun lc ln men ng mannitol cy chuyn sang cc mi trng kim tra cc tnh cht sinh ho hc ca t cu. Tiu chun chn on t cu vng: + Cu khun hnh chm nho bt mu Gram(+). + Khun lc dng S, mu vng, tan mu. + Ln men ng mannitol. + Coagulase(+), Catalase(+). 1.1.5. Phng v iu tr bnh Phng bnh: Vaccin phng bnh t cu t c kt qu. Phng php phng bnh ch yu l v sinh mi trng, v sinh c nhn, v sinh n ung. c bit l trnh nhim khun bnh vin, i vi cc dng c tim truyn, cc dng c dng trong sn khoa, ngoi khoa phi m bo v khun trc khi dng cho bnh nhn. iu tr: T cu b tiu dit bi nhiu khng sinh nh: oxacillin, kanamycin, gentamycin,... Tuy nhin, do vic dng khng sinh rng ri v tu tin nn t cu vng khng li nhiu loi khng sinh. Vic iu tr cn thit phi da vo khng sinh chn thuc thch hp.
1.2. Staphylococcus epidermidis v Staphylococcus saprophyticus

Hai loi t cu ny l nhng vi khun bnh thng, k sinh trn da v nim mc, c phn bit vi t cu vng bng tnh cht sau: khng c coagulase, khng c sc t vng, khng ln men ng manitol. S k sinh ca vi khun c ngha: Chng li s xm nhp ca cc vi khun khc, do cc vi khun k sinh chim mt receptor l ch bm trn b mt nim mc. Trn c a suy gim min dch, loi t cu ny c th gy bnh c hi, m rt thng gp l: S. epidermidis gy nhim khun da v vim ni tm mc cp. S. saprophyticus gy nhim khun tit niu. Cc vi khun ny c tr trn c th v trong mi trng bnh vin c th truyn cho nhau kh nng khng thuc, lm cho s khng khng sinh tng ln, v vic iu tr cng kh khn.
1.3. Lin cu khun (Streptococcus)

Lin cu c Billroth m t ln u tin vo nm 1874 sau khi phn lp t m cc tn thng vim qung da v cc vt thng nhim trng. Nm 1880, L. Pasteur phn lp c lin cu bnh nhn nhim khun huyt. 1.3.1. c im sinh vt hc 1.3.1.1. Hnh th v tnh cht bt mu Lin cu l nhng cu khun hnh cu ng knh 0,6 1m, xp lin tip vi nhau thnh tng chui. i khi c v, khng c lng, khng sinh nha bo, bt mu Gram(+).
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1.3.1.2. Tnh cht nui cy Lin cu hiu kh, k kh tu tin, mi trng nui cy cn nhiu cht dinh dng nh mu, huyt thanh, ng,... Vi khun pht trin tt hn iu kin kh trng c thm CO2. Nhit thch hp l 37OC, mt s pht trin c 10 40OC nh lin cu ng rut. Trong mi trng lng, lin cu pht trin hnh thnh nhng chui di khng b gy, sau to thnh nhng ht nh, hoc nhng ht nh bng ri lng xung y. Sau 24 gi, mi trng nui cy tr nn trong v c lng cn. Trong mi trng c, vi khun pht trin thnh nhng khun lc trn, li, bng kh, mu hi xm. Trong mi trng thch mu lin cu pht trin tt, c th lm tan mu di 3 hnh thc , , g tu thuc tng nhm lin cu: + Tan mu (): Tan mu khng hon ton, xung quanh khun lc c vng tan mu mu xanh, thng gp lin cu Viridans. + Tan mu (): Tan mu hon ton, xung quanh khun lc c vng tan mu trong sut. Tan mu ch yu lin cu nhm A, ngoi ra c th gp nhm B, C, G, F. + Tan mu (g): xung quanh khun lc khng c vng tan mu, hng cu trong thch vn gi mu hng. Thng gp tan mu kiu ny i vi S. faecalis. 1.3.1.3. Tnh cht sinh ho hc Lin cu khng c men catalase. Lin cu c kh nng pht trin mi trng c mt, mui mt, etyl hydrocuprein. Lin cu nhm A c bit nhy cm vi bacitracin. 1.3.1.4. Cu trc khng nguyn Lin cu c cu trc khng nguyn rt phc tp. Khng nguyn C c hiu nhm: Nm 1930, Lancefield da vo khng nguyn C (carbohydrat) ca vch t bo vi khun xp lin cu thnh cc nhm t A n R. Lin cu nhm A v D c kh nng gy bnh cho ngi. Cc nhm khc gy bnh cho sc vt hoc khng gy bnh. Khng nguyn M (Protein M): c hiu typ nm vch t bo vi khun. Da vo khng nguyn ny Lancefiel xp lin cu nhm A thnh 80 typ huyt thanh khc nhau. Khng nguyn M c kh nng chng li thc bo, v vy, n lin quan trc tip ti c lc ca lin cu.
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Nhng khng nguyn khc ca lin cu: + Khng nguyn T: l protein ca vch t bo vi khun. + Khng nguyn P: bn cht l nucleoprotein. + Khng nguyn R: bn cht l protein, nm vch t bo vi khun. 1.3.1.5. Cc enzym v c t Streptokinase thng lin cu nhm A, C, G, c kh nng lm tan t huyt, hot ho xung quanh vng tn thng, to iu kin cho vi khun lan trn. Enzym ny cn l mt khng nguyn c kh nng kch thch c th hnh thnh khng th antistreptokinase. Ngi ta sn xut khng nguyn ny t lin cu nhm C v ng dng trong iu tr bnh ng fibrin trong mu. Streptodornase c kh nng thu phn ADN, lm lng m nhng ch c tc dng khi c mt ca ion Mg. Streptodornase c 4 loi A, B, C, D v u l nhng khng nguyn kch thch c th hnh thnh khng th c hiu. Hyaluronidase c tc dng phn hu acid hyaluronic ca t chc, to iu kin cho vi khun lan truyn su rng vo cc m. Enzym ny cng c tnh khng nguyn kch thch c th sinh khng th c hiu. DPNase (Diphospho pyridine nucleotidase) c lin cu nhm A, C, G, c c tnh vi t bo bch cu, gy cht bch cu v cng l mt enzym c tnh khng nguyn, kch thch c th to khng th. Proteinase c tc dng thu phn protein v kch thch c th hnh thnh khng th. Dung huyt t lin cu tan mu () c kh nng hnh thnh hai loi dung huyt t. + Streptolysin O b mt hot tnh bi oxy. c t ny mang tnh cht ca mt ngoi c t, c tnh khng nguyn mnh nn kch thch c th hnh thnh khng th anti streptolysin O (ASLO). Vic nh lng khng th ny c gi tr trong chn on bnh do lin cu, c bit l bnh thp tim v vim cu thn cp. + Streptolysin S khng b mt hot tnh bi oxy, tnh khng nguyn yu nn khng kch thch c th hnh thnh khng th. c t hng cu (erythrogennictoxin) cn c gi l c t sinh , bn cht l protein gy pht ban trong bnh tinh hng nhit. 1.3.2. Kh nng gy bnh 1.3.2.1. Gy bnh cho ngi
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a) Bnh do lin cu nhm A Lin cu nhm A gy bnh quan trng nht ngi, tu tng typ huyt thanh m gy nn cc th lm sng: Nhim khun ti ch: Vim hng, eczema, trc l, nhim khun vt thng, vim tai gia, vim phi, nhim trng t cung sau ,... Cc nhim khun th pht: T nhim khun ti ch bnh nhn c th b nhim khun huyt, vim mng trong tim cp. Bnh tinh hng nhit: Bnh thng gp cc nc n i, tr em trn 2 tui d mc bnh hn. Cc biu hin lm sng ng ch nht l thn v tim. Cc bnh khc: + Vim cu thn sau nhim lin cu nhm A: thng do typ 12, v mt s typ 4, 18, 25, 49, 52, 55. Bnh thng xy ra sau khi mc vim hng hoc vim da do lin cu. Cc gi thuyt cho rng s tc ng ca khng th chng li khng nguyn vch ca lin cu nhm A phn ng cho vi mng y ca cu thn. + Bnh thp tim: Bnh thng xy ra sau nhim lin cu nhm A hng 2 3 tun, sau khi c th hnh thnh khng th chng lin cu. Thng gp mt s typ huyt thanh: 1, 3, 5, 6, 14, 18, 19, 24, 27, 29 gy thp tim. Nguyn nhn do phn ng cho ca glycoprotein van tim, bao khp vi khng th chng lin cu nhm A. b) Bnh do lin cu nhm D Lin cu nhm D l mt trong nhng vi khun ch bnh thng ng rut v gy bnh khi gp iu kin thun li. Lin cu nhm D c th gy nhim khun ng tit niu, nhim khun huyt, vim mng no, i khi gy vim mng trong tim. c) Bnh do lin cu Viridans Lin cu Viridans khng tan mu hoc tan mu , gy nhim khun ng h hp v l cn nguyn chnh gy vim mng trong tim chm (osler) trn nhng ngi c cu trc van tim khng bnh thng. 1.3.2.2. Gy bnh thc nghim Th l sc vt cm nhim i vi lin cu. Th c th biu hin cc bnh cnh khc nhau nh apxe, vim khp, nhim khun huyt. 1.3.2.3. Min dch Trong cc loi khng th c to thnh, ch c khng th khng protein M c kh nng chng li qu trnh nhim trng, khng th ny mang tnh c hiu typ. 1.3.3. Chn on vi khun hc 1.3.3.1. Chn on trc tip Ly bnh phm: Bnh phm c th l m vt thng, dch ngoy hng, mu, nc, no tu, dch apxe,... tu tng th bnh. Cc loi bnh phm phi c nui cy ngay, hoc chm nht khng qu 3 gi. i vi mu phi ly vo lc bnh nhn ang st, trng hp chn on vim mng trong tim phi ly mu nhiu ln. Nhum soi: Bng phng php nhum Gram, nu thy cu khun Gram(+) xp thnh chui th tip tc phn lp xc nh lin cu. Nui cp phn lp: + Bnh phm l mu, dch no tu: Nui cy vo bnh canh thang glucose, 37OC, theo di hng ngy. Nu mi trng trong sut, y c lng cn th nhum soi, khi thy lin cu th tip tc cy chuyn v kim tra cc tnh cht sinh ho hc. + Bnh phm l m, dch ngoy hng: Nui cy bnh phm vo mi trng thch mu, t m 37OC, theo di s hnh thnh khun lc v tnh cht tan mu. Nhum soi li hnh th, nu l cu khun Gram(+) ng thnh chui th xc nh tnh cht sinh ho hc. c bit xc nh tnh cht lin cu nhm A bng th nghim bacitracin. Phn bit lin cu vi ph cu bng th nghim Optochin v Neufeld.
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Hin nay cn c phng php chn on nhanh lin cu nhm A t tm bng ngoy hng bng cch s dng phn ng ngng kt latex, y l phng php c c hiu cao nhng nhy km. Tiu chun chn on lin cu: + Hnh th v tnh cht bt mu. + Hnh thi khun lc v tnh cht tan mu trn mi trng thch mu. + Lin cu nhm A nhy cm vi bacitracin. + Optochin hoc Neufeld (). + Catalase (). 1.3.3.2. Chn on gin tip Tm khng th trong huyt thanh chn on bnh nhn mc bnh do lin cu nhm A. c bit, xt nghim anti streptolyzin O (ASLO) l xt nghim c s dng trong chn on bnh thp tim v vim cu thn cp tr em. 1.3.4. Phng bnh v iu tr Phng bnh: Hin nay cha c vaccin phng bnh, nn ch yu vn l v sinh c nhn, v sinh mi trng. trnh nhim trng th pht v bin chng khp, tim, vim cu thn th cn phi pht hin sm v iu tr tch cc cc nhim khun da, hng do lin cu nhm A. S dng khng sinh thch hp phng lin cu sau cc phu thut ng h hp, tit niu,... iu tr: Cc khng sinh penicillin, ampicillin,... vn c tc dng vi lin cu nhm A. Lin cu Viridans, lin cu ng rut khng khng sinh mnh, do iu tr phi da vo khng sinh .
1.4. Ph cu khun (Steptococcus pneumoniae)

Ph cu c phn lp ln u bi L. Pasteur Php nm 1880. 1.4.1. c im sinh vt hc 1.4.1.1. Hnh th v tnh cht bt mu Ph cu khun l nhng song cu hnh ngn nn, hai u to gip vo nhau, hai u nhn quay ra ngoi. Trong mi trng nui cy c th nhiu i xp lin tip vi nhau thnh tng chui d lm vi lin cu. Bt mu Gram(+), khng di ng, khng sinh nha bo, trong bnh phm hay trong mi trng nui cy c nhiu albumin th c v.
1.4.1.2. Tnh cht nui cy Ph cu pht trin tt mi trng c nhiu cht dinh dng, kh trng c 5% CO2, nhit thch hp 37OC. Trn mi trng thch mu, khun lc nh, trn, li, bng, trong nh git sng, xung quanh c vng tan mu . Nhng ph cu c v khun lc thng ln, hi nhy, mu xm nht.
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1.4.1.3. Tnh cht sinh ho hc Ph cu khng c men catalase. B ly gii bi mt hoc mui mt (th nghim Neufeld dng). Khng pht trin c mi trng c etyl hydrocuprein (th nghim Optochin +). 1.4.1.4. Cu to khng nguyn Khng nguyn v: V ca ph cu c cu to bi polysaccharid c tnh khng nguyn c hiu typ. Hin nay c 85 typ huyt thanh ca ph cu c ghi nhn bi khng nguyn ny. Khng nguyn thn: Ph cu c 3 loi khng nguyn thn: khng nguyn polysaccharid C l khng nguyn c hiu loi, khng nguyn M l nhng protein c hiu typ v khng nguyn R hiu bit cn t. 1.4.1.5. Sc khng Ph cu d b tiu dit bi nhit (60OC/30 pht) v cc thuc st khun thng thng. 1.4.1.6. Cc yu t c lc Ph cu khng c ni v ngoi c t. Ph cu gy bnh ch yu do v ca vi khun c tc dng v hiu ho khng th IgG v b th. Do vy, kh nng thc bo b gim xung v vi khun vn tn ti gy bnh. Ph cu cn tit protease thu phn IgA tit, v vy, lm mt tc dng ngn cn s xm nhp ca vi khun vo nim mc h hp. 1.4. 2. Kh nng gy bnh 1.4.2.1. Gy bnh cho ngi Ph cu sng t hu ngi lnh vi t l cao 40 70%. Ph cu gy nn cc bnh ng h hp, in hnh l vim mi hng, vim phi, vim ph qun phi, apxe phi, vim mng phi. Vim phi do ph cu thng xy ra sau cc bnh nhim khun ng h hp do virus nh cm hoc do ho cht. Cc typ gy bnh thng l typ 1, 2, 3 i vi ngi ln v 4, 1, 6 i vi tr em. Ngoi ra, ph cu cn gy vim tai, vim xoang, vim mng no, vim phc mc, nhim khun huyt, vim mng tim, vim thn,... ni tn thng, ph cu hnh thnh mt lp v dy v c nhiu fibrin bao quanh to nn mt vng cch bit lm cho khng sinh kh tc dng. V vy, nn iu tr sm v trit . 1.4.2.2. Gy bnh thc nghim Sc vt cm nhim l chut nht trng, chut bch, th, kh. 1.4.3. Chn on vi khun hc 1.4.3.1. Chn on trc tip Bnh phm: Cht ngoy hng, m, mu, dch no tu, dch mng phi, m,... tu theo tng tn thng. Ph cu l vi khun rt d cht khi ra ngoi mi trng, v vy bnh phm phi c chuyn v phng xt nghim trong vng 2 4 gi v gi 4 8OC. Nhum soi: Nhum Gram, t c gi tr trong trng hp bnh phm l cht ngoy hng, dch hng mi hoc m. Nui cy phn lp: + Bnh phm c nhim nhiu vi khun nh m, cht ngoy hng c cy vo mi trng thch mu c gentamycin 5g1ml mi trng. iu kin thch hp, nhng ph cu c v khun lc s c nh, sau 24 gi khun lc lm gia. Tnh cht ny gip phn bit vi lin cu viridans. + Bnh phm l mu, dch no tu c cy vo mi trng canh thang mu, t m 37OC c 5% CO2, theo di nh phn lp lin cu. Cc th nghim xc nh ph cu:
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+ Th nghim Neufeld. + Th nghim Optochin. + Xc nh c lc ca ph cu gy bnh bng thc nghim trn chut nht trng. + Xc nh v ca vi khun bng phng php nhum v hoc dng phng php phnh v (Quellung). 1.4.3.2. Chn on gin tip Chn on gin tip khng c ngha trong chn on ph cu nn khng c dng trong phng xt nghim. Tiu chun chn on ph cu: Hnh th v tnh cht bt mu. Hnh thi khun lc trn mi trng thch mu c gentamicin. B tan bi mui mt hoc sc t mt. Gy bnh cho sc vt. 1.4.4. Phng bnh v iu tr Phng bnh: Vaccin phng bnh hin nay c s dng cc nc tin tin, tc dng bo v ca vaccin khng cao, nhng c tc dng ngn cn nhim ph cu nng nh nhim khun huyt, vim mng no m v vaccin khng y cc serotyp. Ph cu ly theo ng h hp nn vic phng bnh khng c hiu rt kh khn. Ch yu v sinh ng h hp. iu tr: Ph cu cn nhy cm vi cc khng sinh thng dng nh penicillin, gentamycin, cephalosporin.
NEISSERIA
Neisseria l nhng cu khun Gram(), khng di ng, khng c v, thng xp thnh tng i, c chia thnh hai nhm: Nhm Neisseria phi phn lp trn mi trng c bit, nhm ny khng sinh sc t. C hai loi vi khun gy bnh cho ngi l N.menigitidis (no m cu) v N.gonorrhoeae (lu cu) Nhm cc Neisseria phn lp c trn mi trng nui cy thng thng, nhm ny c sc t, gm 14 loi, k sinh ng h hp trn v rt t khi gy bnh.
1.5. No m cu (Neisseria meningitidis)

Neisseria meningitidis c Albrecht v Ghon m t ln u tin vo nm 1901 v t tn vo nm 1903. 1.5.1. c im sinh vt hc 1.5.1.1. Hnh th v tnh cht bt mu No m cu l loi song cu hnh ht c ph hai mt lm quay vo nhau, trng ging nh hnh ht c ph, kch thc khong 1m, ng ring r tng i hoc nhiu i t vi nhau thnh tng m. C th nm trong hoc ngoi bch cu a nhn, khng lng, khng di ng, khng sinh nha bo, bt mu Gram().
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1.5.1.2. Tnh cht nui cy No m cu ch pht trin tt mi trng giu cht dinh dng nh thch mu, thch chocolat, kh trng c 5 10% CO2. Nhit thch hp l 37OC. Trn mi trng thch mu, khun lc trn, nhn, li, bng, sau 24 gi ng knh khong 1mm, khng tan mu. Trn mi trng thch chocolat, khun lc dng S, xm hoc ng nh. 1.5.1.3. Tnh cht sinh ho hc Ln men khng sinh hi cc loi ng maltose, glucose. Tnh cht ln men ng maltose phn bit vi lu cu. Oxydase (+). 1.5.1.4. Cu trc khng nguyn No m cu c khng nguyn v l polysaccharid. Da vo khng nguyn ny hin nay c 13 nhm c bit, trong c 9 nhm thng gp l A, B, C, D, X, Y, Z,W135, E29, 4 nhm cn li l H, I, K, L him gp hn. Cc nhm A, B, C thng gy bnh thnh dch. Cc khng nguyn v polysaccharid ca no m cu c tm thy trong dch no tu v mu. C th pht hin nhanh khng nguyn bng k thut min dch. Da vo protein mng ngoi t bo li chia mi nhm khng nguyn thnh cc typ huyt thanh. 1.5.1.5. c t No m cu c ni c t vng bn vi nhit . 1.5.1.6. Sc khng No m cu c sc khng yu, d b tiu dit bi cc cht st khun thng thng v iu kin kh, nng v nh sng mt tri. B cht nhit 60OC/10 pht v sau khi ra khi c th 3 4 gi. 1.5.2. Kh nng gy bnh No m cu ch k sinh ngi v gy bnh cho ngi. Chng thng k sinh hng mi ngi bnh thng vi t l 2 8% v khng gy bnh. Khi c iu kin thun li, no m cu gy vim hng mi nhng thng nh, khng c triu chng. Mt t l nh trong cc trng hp ny vi khun t mi hng xm nhp vo mu gy nhim khun huyt. Nhim khun huyt do no m cu c th dn n tnh trng shock do ni c t rt nng. Cng c th vi khun qua mu n mng no gy vim mng no m vi cc triu chng xut hin t ngt nh: nhc u d di, nn, c cng, st cao, hn m. Him hn, c th gp no m cu gy cc tn thng khp v phi.
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1.5.3. Chn on vi khun hc Bnh phm: Dch mi hng, dch no tu, mu, tu theo th bnh. Nhum soi: Nhum soi t dch no tu thy c vi khun trong t bo bch cu a nhn th chn on l nguyn nhn gy bnh. Nu thy vi khun dch mi hng th cn nui cy phn lp phn bit vi cc Neisseria khc. Nui cy: Cy dch no tu vo mi trng thch mu hoc chocolat, iu kin kh trng c CO2 v 37OC, sau 24 gi chn khun lc nghi ng xc nh tnh cht sinh ho hc v lm phn ng ngng kt trn lam knh vi khng th mu xc nh nhm, typ huyt thanh. Tm khng nguyn: ly dch no tu lm phn ng ngng kt vi khng th c hiu c gn trn cc ht latex, k thut ny c th chn on nhanh s hin din ca no m cu trong dch no tu. y l k thut c gi tr cao trong chn on bnh vim mng no m do no m cu. 1.5.4. Phng bnh v iu tr Phng bnh: + Phng c hiu: Hin nay c vaccin ch t v polysaccharid ca no m cu. Vaccin gm 4 nhm khng nguyn (A, C, Y v W135). + Phng khng c hiu: Pht hin sm bnh nhn v cch ly ngay v bnh vim mng no m do no m cu ly bng ng h hp. Dng khng sinh phng cho nhng ngi tip xc vi bnh nhn hoc trong vng dch, thng dng rifampicin hoc minocyclin. iu tr: Nn iu tr sm cho bnh nhn bng penicilin.
1.6. Lu cu (Neisseria gonorrhoeae)

Nm 1879, Neisser l ngi u tin m t vi khun lu l cn nguyn gy bnh lu. Nm 1882, Lestikow v Loeffler nui cy thnh cng lu cu. 1.6.1. c im sinh vt hc 1.6.1.1. Hnh th v tnh cht bt mu Lu cu l loi song cu hnh ht c ph, bt mu Gram(). Trng hp lu cp vi khun ng trong t bo bch cu a nhn trung tnh. Trng hp lu mn tnh, trn tiu bn t vi khun v thng thy vi khun nm ngoi t bo bch cu. Lu cu khng c v, khng c lng, khng sinh nha bo.
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Hnh 8.6. Neisseria gonorrhoeae
1.6.1.2. Tnh cht nui cy Nui cy cu khun lu thng kh khn, sau khi ra khi c th vi khun rt d cht. Mi trng nui cy cu khun lu phi giu cht dinh dng nh mu, huyt thanh v cc yu t pht trin. Kh trng phi c 3 10% CO2 v nhit 35 37OC, m 70%. Trn cc mi trng nui cy nh thch chocolat, ThayerMartin. Sau 24 gi, khun lc mu trng xm, li, trn, lp lnh sng. 1.6.1.3. Tnh cht sinh ho hc Oxydase(+). Catalase(+). Ln men ng glucose. Khng ln men ng maltose, levulose. Da vo s ln men 2 loi ng ny phn bit lu cu vi no m cu khun.
1.6.1.4. Sc khng Cu khun lu c sc khng yu. nhit 58OC vi khun cht sau 5 pht. Sau khi ra khi c th, vi khun cht sau 1 2 gi. Vi cc cht st khun thng thng, vi khun cht sau 2 5 pht tip xc. 1.6.2. Kh nng gy bnh Lu cu c vt ch duy nht l ngi. Bnh ly truyn ch yu bng ng tnh dc. nam, vi khun gy vim niu o, triu chng ch yu l i but, i kh, i m, c th gp vim tin lit tuyn, vim mo tinh hon. ph n, triu chng phc tp, vim niu o, m o, vim c t cung, i khi vim t cung, vi trng, bung trng, cng c khi triu chng khng r rng v kh chn on. C th gp bnh lu tr em, thng gp l vim m kt mc mt sau 1 7 ngy do vi khun ly t ng sinh dc ca ngi m b bnh. Ngoi ra, c th gp nhim cu khun lu lan to, thng gp nhng ngi b bnh nhng khng c iu tr, s c biu hin vim khp, vim gan, vim mng no,... Bnh lu khng c min dch do khng th khng c vai tr bo v. C th chn on huyt thanh trong trng hp bnh lu ngoi ng sinh dc nh vim khp. 1.6.3. Chn on vi khun hc 1.6.3.1. Chn on trc tip Bnh phm + nam: Ly m niu o vo sng sm trc khi i tiu. + n: Ly m niu o, m c t cung hoc cc l ca tuyn m o. Nhum soi: Bnh phm c nhum Gram hoc xanh methylen. Nui cy: Bnh phm c cy vo cc mi trng thch hp. Sau 48 gi, nhn xt hnh thi khun lc, nhum soi li. Xc nh l lu cu bng cc phn ng sinh ho hc. 1.6.3.2. Chn on gin tip
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Trong mt s bnh, nht l vim khp do lu, kt qu nui cy thng m tnh, c th lm cc phn ng huyt thanh chn on. 1.6.4. Phng bnh v iu tr Phng bnh: Vaccin phng bnh khng c hiu qu, ch yu l gii quyt nn mi dm, tuyn truyn gio dc cc bin php phng bnh trong quan h tnh dc. Pht hin sm v iu tr trit cho bnh nhn. c bit, pht hin v iu tr cho ph n c thai b bnh lu, trnh ly sang tr s sinh. iu tr: Hin nay xut hin nhiu chng cu khun lu khng khng sinh, do cn lm khng sinh la chn khng sinh thch hp. Cn iu tr trit khng bnh chuyn sang mn tnh, rt kh chn on v iu tr.
2. H VI KHUN NG RUT (Enterobacteriaceae)

H vi khun ng rut bao gm cc trc khun Gram() c chung nhng tnh cht sau: Di ng hay khng di ng, nu di dng th c lng xung quanh thn. Hiu kh hoc k kh tu tin. Ln men ng glucose c km theo sinh hi hoc khng. Kh nitrat thnh nitrit. Khng c men oxydase. Mc d dng trn cc mi trng nui cy thng thng. Khng sinh nha bo. Vi nhng tnh cht trn, mt s vi khun mc d sng v gy bnh ng tiu ho cng khng c xp vo h vi khun ng rut nh phy khun t. Cc vi khun ng rut gy bnh quan trng l: Trc khun Salmonella. Trc khun Shigella. Trc khun E. coli.
2.1. Trc khun Salmonella

Trc khun thng hn c Grafky phn lp nm 1884. 2.1.1. c im sinh vt hc 2.1.1.1. Hnh th v tnh cht bt mu Trc khun Salmonella c kch thc trung bnh di 3m, ng knh 0,5m, Gram(), c nhiu lng xung quanh thn, rt di ng, khng c v, khng sinh nha bo.
Hnh 8.7. Trc khun Salmonella (nh knh hin vi in t).
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2.1.1.2. Tnh cht nui cy Vi khun hiu kh, k kh tu tin, pht trin d dng trn cc mi trng nui cy thng thng, nhit thch hp l 37OC, pH l 7,6. Trn mi trng lng: sau 5 6 gi nui cy vi khun lm c nh, sau 18 gi mi trng c u. Trn mi trng thch thng khun lc trn, li, bng, thng khng mu hoc mu trng xm. Trn mi trng phn lp c cht c ch chn lc nh SS, Istrati, khun lc c cng mu vi mi trng. 2.1.1.3. Tnh cht sinh ho hc Ln men ng glucose km theo sinh hi (tr S.typhi). Khng ln men ng lactose. Sinh H2S. Catalase (+). Indol (); urease () 2.1.1.4. Cu trc khng nguyn Khng nguyn O: l khng nguyn thn ca vi khun. Da vo khng nguyn O chia Salmonella thnh cc nhm A, B, C, D, E. Khng nguyn H: l khng nguyn lng ca Salmonella. Khng nguyn K: l khng nguyn b mt ca vi khun, ch c S. typhi v S. paratyphi C c gi l khng nguyn Vi (Virulence). 2.1.1.5. c t Ni c t c vai tr quyt nh trong tnh cht gy bnh. 2.1.1.6. Sc khng Trc khun Salmonella b cht nhit 100OC/5 pht. Nhy vi cc thuc st khun thng thng. Trong nc, trc khun sng c 2 3 tun, trong nc v trong phn sng c 2 3 thng. 2.1.1.7. Phn loi Da theo cu trc khng nguyn: S. typhi (trc khun thng hn): Ch gy bnh cho ngi, l cn nguyn quan trng gy bnh thng hn. S. paratyphi A (trc khun ph thng hn A): Gy bnh cho ngi. Vit Nam, l cn nguyn th 2 gy bnh thng hn sau S.typhi. S. paratyphi B (trc khun ph thng hn B): Ch yu gy bnh cho ngi nhng cng c th gy bnh cho ng vt. S. paratyphi C (trc khun ph thng hn C): gy bnh thng hn, gy vim d dy rut v nhim khun huyt. Thng gp cc nc ng Nam . Ngoi 4 loi Salmonella trn, cn c mt s Salmonella khc c t tn theo vt ch hoc theo tn a phng phn lp c nh: S. typhimurium (vt ch l chut) v S. enteritidis: C kh gy bnh cho c ngi v ng vt, l cn nguyn ch yu gy vim d dy rut cp. n nay, pht hin c trn 1.500 typ huyt thanh Samonella. 2.1.2. Kh nng gy bnh 2.1.2.1. Gy bnh thng hn Cc Salmonella gy bnh bao gm S. typhi v S. paratyphi A, B, C. C ch gy bnh: Vi khun xm nhp vo c th theo ng tiu ho do thc n, nc ung b
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nhim bn. S lng vi khun gy bnh khong 105 n 107. Sau khi vo ng tiu ho, vi khun bm vo nim mc rut non, xm nhp qua nim mc rut n cc hch mc treo, sinh sn nhn ln y ri qua h thng bch huyt v ng ngc vo mu, lc ny cc triu chng bnh bt u xut hin. T mu, vi khun qua lch v cc c quan khc. Khi qua gan, vi khun theo ng dn mt xung rut ri c o thi ra ngoi theo phn, y l ng thi vi khun chnh. Hoc t mu, vi khun n thn v c o thi ra ngoi nc tiu. Ti mng payer rut, vi khun tip tc nhn ln. Trc khun thng hn gy bnh bng ni c t. Ni c t kch thch thn kinh giao cm rut gy hoi t, chy mu v c th gy thng rut. Ni c t theo mu ln kch thch trung tm thn kinh no, bnh nhn thng st cao, thn nhit tng nhng nhp tim khng tng (mch nhit phn ly), tnh trng ly b, c th hn m, tru tim mch, t vong. Khong 5% bnh nhn sau khi khi bnh tr thnh ngi lnh mang trng, y l ngun truyn bnh rt nguy him v vn tip tc thi vi khun qua phn do ngun vi khun vn tn ti ti mt. 2.1.2.2. Gy nhim khun nhim c thc n Nhim khun nhim c thc n hay vim d dy rut cp, thng gp do S. typhymurium, S. enteritidis. Sau khi n phi thc n b nhim khun 10 48 gi, bnh nhn c biu hin st, nn, tiu chy, c th dn n mt nc in gii nu khng kp thi iu tr. Trong trng hp ny, vi khun ch gy bnh ti ng tiu ho, khng xm nhp vo bch huyt v mu. 2.1.2.3. Min dch Sau khi mc bnh thng hn, trong huyt thanh bnh nhn c cc khng th chng khng nguyn O, H v Vi. Khng th IgA trong dch tit ng rut c vai tr quan trng trong c ch bo v. Tuy nhin, vai tr bo v ca khng th khng y . 2.1.3. Chn on vi khun hc 2.1.3.1. Chn on trc tip Cy mu: Cy mu vo lc bnh nhn ang st v cha iu tr khng sinh. T l dng tnh khi cy mu tun u ti 90%, tun th 2 t 70 80%, tun th 3 t 40 60%. Cy mu c vi khun cho php chn on chc chn bnh nhn mc bnh thng hn. Cy phn: V trong phn c nhiu tp khun nn nui cy vo mi trng phn lp thch hp. Sau 24 gi, chn khun lc khng ln men ng lactose cy chuyn sang cc mi trng xc nh tnh cht sinh ho hc. 2.1.3.2. Chn on gin tip Tm khng th trong huyt thanh bng phn ng ngng kt Widal. Kt qu ln xt nghim th nht thng khng cho php xc nh chc chn. Phn ng cn c lm 2 ln, tun th nht v th hai xc nh ng lc khng th. Nu ng lc khng th cao mi cho php chn on. 2.1.4. Phng bnh v iu tr Phng bnh: + Phng c hiu bng vaccin. + Phng khng c hiu l cc bin php nh v sinh thc phm, v sinh mi trng, pht hin sm bnh nhn v cch ly kp thi. iu tr: T l vi khun khng khng sinh ngy cng tng nn iu tr tt nht l theo khng sinh .
2.2. Trc khun Shigella
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Trc khun Shigella c Chantemesse m t t nm 1888 v Shiga phn lp ln u tin nm 1898. 2.2.1. c im sinh vt hc 2.2.1.1. Hnh th v tnh cht bt mu Shigella l nhng trc khun di 1 3m, bt mu Gram(), khng c lng, khng c v, khng sinh nha bo.
Hnh 8.8. Trc khun Shigella
2.2.1.2. Tnh cht nui cy Vi khun hiu kh, k kh tu tin, pht trin d dng trn cc mi trng nui cy thng thng. Nhit thch hp l 37OC, pH l 7,8. Trong mi trng lng, vi khun mc sm v lm c u mi trng. Trn mi trng c, khun lc trn, li, b u. Trn mi trng phn lp c ng lactose nh Istrati, SS: khun lc c mu cng vi mu mi trng. 2.2.1.3. Tnh cht sinh ho hc Ln men ng glucose khng sinh hi (tr S. flexneri 6, S. Boydii 14 sinh hi yu). Khng ln men ng lactose (tr S. sonnei ln men chm sau 2 ngy n 2 tun). Ln men ng malnitol (tr S. dysenteriae). Shigella khng sinh H2S v indol. 2.2.1.4. Cu trc khng nguyn Tt c cc Shigella u c khng nguyn thn O, y l khng nguyn quan trng nht, mt s chng c khng nguyn b mt K v tt c u khng c khng nguyn H. 2.2.1.5. c t Cc Shigella u c ni c t v mt s chng c thm ngoi c t. Ni c t: C c tnh mnh. Ngoi c t: Ch c chng S. schmitzii v S. shiga. Ngoi c t c c tnh cao, tc dng c hiu vo h thn kinh trung ng. 2.2.1.6. Sc khng Shigella tn ti c trong nc v thc n 7 10 ngy, trong t 6 7 tun. Tuy nhin, vi khun b cht nhanh nc si 100OC, b tiu dit bi cc thuc st khun thng thng v nh sng mt tri.
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2.2.2. Phn loi Da vo khng nguyn thn O v tnh cht sinh ho hc, cc trc khun Shigella c chia lm 4 nhm. Nhm A (S. dysenteriae) c 10 typ huyt thanh. ng ch l typ 1 (S. shiga) v typ 2 (S. schmitzii) ngoi ni c t cn sinh ngoi c t. Nhm B (S. flexneri) c 13 typ huyt thanh. Nhm C (S. boydii) c 15 typ huyt thanh. Nhm D (S. sonnei) ch c 1 typ huyt thanh. 2.2.3. Kh nng gy bnh Trc khun Shigella gy nn bnh l ngi. Trc khun xm nhp vo c th bng ng n ung, n c tr v sinh sn rt nhanh nim mc i trng. Vi khun gy bnh nh kh nng xm nhp v ni c t. Ni c t gy sung huyt, xut tit to thnh nhng lot v mng hoi t nim mc i trng; tc ng ln thn kinh giao cm gy co tht v tng nhu ng rut. Bnh biu hin bng hi chng l: au qun, mt rn, phn c nhy mu. Ngoi c t ca trc khun Shigella c tnh c vi thn kinh trung ng, c th gy vim mng no v hn m. Bnh l trc khun thng th cp tnh. Mt s t trng hp tr thnh mn tnh, nhng bnh nhn ny thnh thong li c tiu chy v thi vi khun ra ngoi theo phn. Bnh do S. shiga thng gy nn nhng v dch ln v ko di, nng hn cc chng khc. Vit Nam hin nay, a s cc trng hp l trc khun l do nhm B v C. Sau khi mc bnh l trc khun, c th c khng th c hiu, tuy nhin, hiu lc bo v ca cc khng th ny rt km. Vai tr bo v ch yu l nh IgA tit ti rut. 2.2.4. Chn on vi khun hc 2.2.4.1. Chn on trc tip Bnh phm: Ly bnh phm l phn. Nhum soi trc tip: Lm tiu bn nhum soi xc nh mt bch cu a nhn v vi khun ch. Cy phn: Cy phn l phng php c dng ch yu chn on bnh l trc khun. Cy phn cng sm cng tt v trc khun l cht nhanh sau khi ra khi c th. Mi trng phn lp thng dng l SS, Endo, Istrati. Sau 24 gi, chn khun lc nghi ng xc nh tnh cht sinh ho hc. Phn ng ngng kt xc nh nhm v typ huyt thanh. Sau khi nui cy xc nh l Shigella th tin hnh xc nh nhm bng khng huyt thanh a gi nhm. Nu mt trong cc khng huyt thanh a gi nhm ngng kt th tin hnh phn ng ngng kt vi cc khng huyt thanh n gi theo tng nhm. 2.2.4.2. Chn on huyt thanh Phn ng huyt thanh rt t c lm chn on bnh l trc khun, v y l bnh cp tnh cn chn on nhanh. 2.2.5. Phng bnh v iu tr Phng bnh: + Hin nay cha c vaccin phng bnh. + Thc hin cc bin php phng bnh khng c hiu: V sinh an ton thc phm, v sinh mi trng, pht hin sm, cch ly bnh nhn. iu tr: Shigella l mt trong s cc vi khun c t l khng khng sinh rt cao. V vy, cn phi lm khng sinh chn khng sinh thch hp.
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2.3. Trc khun Escherichia coli

2.3.1. c im sinh vt hc 2.3.1.1. Hnh th v tnh cht bt mu Trc khun E. coli kch thc di, ngn khc nhau, trung bnh di 2 3m, ng knh 0,5m, Gram(). Mt s t chng c v, hu ht c lng v di ng, khng sinh nha bo.
Hnh 8.9. Trc khun Escherichia coli (nh di knh hin vi in t)
2.3.1.2. Tnh cht nui cy E. coli hiu kh, k kh tu tin, pht trin d dng trn cc mi trng nui cy thng thng. Nhit thch hp l 37OC. Trong iu kin thch hp, E. coli pht trin nhanh, thi gian th h ch khong 20 n 30 pht. Trong mi trng lng, sau 4 5 gi nui cy, vi khun lm c nh, sau 24 gi mi trng c u, sau 2 ngy mi trng c vng mng. lu vi khun lng xung y ng nghim. Trn mi trng thch thng, sau 18 24 gi khun lc dng S, trn, b u, li bng, khng mu hoc mu xm nh. Trn mi trng phn lp, vi khun thng lm thay i mu mi trng v ln men ng lactose, khun lc c mu vng trn mi trng Istrati, mu trn mi trng SS. 2.3.1.3. Tnh cht sinh ho hc E. coli ln men nhiu loi ng v sinh hi nh glucose, malnitol, lactose (tr loi EIEC lactose ()). E. coli c kh nng sinh indol. Khng sinh H2S. Khng c urease. 2.3.1.4. Cu trc khng nguyn Khng nguyn O l khng nguyn thn gm 160 yu t. Khng nguyn K l khng nguyn b mt. C khong 100 yu t, c chia lm 3 loi A, B v L. Khng nguyn H l khng nguyn lng, gm hn 50 yu t. 2.3.1.5. Sc khng Nhy cm vi cc thuc st khun thng thng. nhit 55OC, vi khun cht sau 1 gi, 60OC cht sau 30 pht. 2.3.2. Phn loi Da vo cu trc khng nguyn, E. coli c chia thnh nhiu typ huyt thanh. C nhiu typ huyt thanh khc nhau, mi typ huyt thanh c k hiu bng khng nguyn O v K, v d: 0111B4
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(yu t khng nguyn O s 111, yu t khng nguyn K s 4 loi B). Da vo tnh cht gy bnh, E. coli c chia thnh cc loi: Nhm E. coli gy bnh ng rut EPEC (Entero pathogenic E. coli). Nhm E. coli sinh c t rut ETEC (Entero toxigenic E. coli). Nhm E. coli xm nhp (Enteroinvasive E. coli). Nhm E. coli bm dnh ng rut EAEC (Enteroadherent E. coli). Nhm E. coli gy chy mu ng rut EHEC (Entero haemorrgagic E. coli). 2.3.3. Kh nng gy bnh E. coli l vi khun bnh thng rut ngi, c bit i trng, chim t l cao nht trong s cc vi khun hiu kh (80%). E. coli l vi khun gy bnh quan trng, ng hng u trong cc vi khun gy tiu chy, vim ng tit niu, vim ng mt. ng hng th hai sau t cu khun trong nhim khun huyt. E. coli cn gy nhiu bnh khc nh vim phi, vim mng no, nhim khun vt thng, vim phc mc c bit sau thng rut. E. coli gy tiu chy cp tr em nht l tr em di 2 tui, bnh c tnh cht dch v gy t vong cao tr em. Mt s tnh cht gy bnh ca cc nhm nh sau: Nhm ETEC: Gy bnh do ngoi c t LT (Labiletoxin). LT l loi c t rut ging c t ca phy khun t. Loi c t ny bm vo th th rut lm gim hp thu Na+, tng tit nc v ion Cl dn n tiu chy cp mt nc v in gii. Nhm EIEC: Gy bnh do c kh nng xm nhp vo nim mc i trng, gy bnh ging trc khun l. Nhm EAEC: Gy bnh do bm vo nim mc lm tn thng chc nng rut. Nhm EHEC: Lm tn thng xut huyt rut, nhm ny c c t c cu trc khng nguyn v c ch tc ng ging ngoi c t ca S. shiga. Nhm EPEC: C ch gy bnh cha r. 2.3.4. Chn on vi khun hc 2.3.4.1. Chn on trc tip Bnh phm: Bnh phm khc nhau tu theo th bnh, c th l mu, phn, nc tiu, m, dch tit. Nhum soi trc tip. Nui cy phn lp: + Bnh phm l mu: Cy vo mi trng canh thang, theo di hng ngy. Nu mi trng c th nhum soi, nu l trc khun Gram() th xc nh cc tnh cht sinh ho hc. + Bnh phm l phn, dch, m: Cy vo mi trng chn lc nh Endo, Macconkey, DCl, Istrati, sau 18 24 gi, chn khun lc nghi ng xc nh cc tnh cht sinh ho hc ca trc khun E. coli. + Nu bnh phm l nc tiu th ly nc tiu gia dng, tin hnh cy, m trn mi trng thch thng. Phn ng ngng kt: Lm phn ng ngng kt trn lam knh vi khng huyt thanh a gi v n gi xc nh typ huyt thanh. 2.3.4.2. Chn on gin tip: Khng s dng chn on. 2.3.5. Phng bnh v iu tr Phng bnh: Hin nay cha c vaccin phng bnh c hiu. Cn ch v sinh n ung, nht l khi c dch vim d dy rut tr em. Thc hin nghim tc nguyn tc v khun khi tin hnh thm d hoc t thng ng tit niu.
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iu tr: E. coli l vi khun c t l khng thuc cao, nht l cc chng phn lp c t nc tiu. V vy, cn lm khng sinh chn khng sinh thch hp trong iu tr. Trong trng hp tiu chy, ngoi vic s dng khng sinh, phi ch bi ph nc in gii cho bnh nhn.
3. PHY KHUN T (Vibrio cholerae)

Vibrio cholerae c R. Koch pht hin nm 1883.
3.1. c im sinh vt hc
3.1.1. Hnh th v tnh cht bt mu Phy khun t c hnh hi cong nh du phy, bt mu Gram(). Vi khun c lng mt u, di ng rt nhanh v mnh, khng c v, khng sinh nha bo
Hnh 8.10. Vibrio cholerae (nh di knh hin vi in t)
3.1.2. Tnh cht nui cy Phy khun t hiu kh, mi trng thch hp c pH kim 8,5 9,5 v nng mui cao 3%, nhit 37OC. Trong mi trng pepton kim vi khun mc nhanh, sau 6 8 gi to thnh vng. Trn mi trng thch kim sau 18 gi khun lc trn, li, nhn v trong sut. Trn mi trng TCBS (Thiosulfat, Citrate Bile Salts, Saccarose). Sau 18 gi khun lc trn, bng, mu vng (ln men ng saccarose). 3.1.3. Tnh cht sinh ho hc Ln men khng sinh hi ng glucose, saccarose, manose. Khng ln men ng lactose, arabinose. Oxydase (+), indol (+). H2S (), urease (). 3.1.4. Cu to khng nguyn v phn loi Phy khun t c 2 loi khng nguyn: Khng nguyn H l khng nguyn chung cho tt c cc loi vi khun t, d b ph hu O 100 C/2 gi. Khng nguyn O l mt lipopolysaccharid, c tnh c hiu cao, b ph hu 100OC/1 gi. Cn c vo s khc nhau ca khng nguyn O, Vibrio cholerae c chia thnh hn 100 nhm, cc chng gy dch t trc nm 1992 u thuc nhm O1. Nm 1992 xut hin mt nhm mi l O139
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gy dch t nhiu nc trn th gii. Da vo cu trc khng nguyn chia Vibrio cholerae O1 ra 3 typ huyt thanh:
Da vo tnh cht sinh hc, vi khun t c chia thnh 2 typ sinh hc (sinh typ): V. cholerae sinh typ c in (gi tt l V. cholerae) c R.Koch phn lp nm 1883 trong mt v dch ln Ai Cp v V. cholerae sinh typ Eltor (gi tt l V. Eltor). Eltor l tn trm cch ly bnh nhn t, ni m Gotschlich phn lp c vi khun t t thi bnh nhn nm 1905. S khc nhau v tnh cht gia 2 typ sinh hc ny l:
3.1.5. Sc khng Phy khun t c sc khng yu, d b tiu dit bi cc cht st khun thng thng, nhng c th sng c mt vi gi trong phn v mt vi ngy trong nc.
3.2. Kh nng v c ch gy bnh

Gy bnh t ngi: + Xm nhp vo c th bng ng n ung. + Thi gian bnh ngn, c th mt vi ngy. + C ch: Vi khun gy bnh khi vt qua d dy ngi c acid gim. (Vi khun d cht khi qua d dy ngi (bnh thng c pH xp x 3). i vi nhng ngi d dy tit dch bnh thng th thc n, nc ung phi c kh nng trung ho bt acid ca dch v, vi khun mi c th gy bnh c). Sau khi xung rut non, vi khun bm vo nim mc nhng khng xm nhp nn khng gy tn thng cu trc nim mc rut. Vi khun pht trin nhanh nh pH thch hp (pH 8), tit ra c t rut LT (thermolabile toxin), LT gn vo th th ph hp trn mng t bo nim mc rut non lm hot ho men adenyl cyclase dn n tng qu nhiu AMP vng, lm cho t bo nim mc rut gim hp thu Na+, tng tit nc v Cl gy tiu chy cp. Bnh nhn cht v mt nc v in gii. Min dch ca bnh: + C ch khng khng c hiu: acid ca dch v, h vi khun ch cnh tranh v tr bm vi phy khun t. + C ch khng c hiu: Bnh c kh nng to min dch kh bn vng, thi gian bo v ca khng th khong 3 nm, vi vai tr quan trng ca IgA tit ng rut. Dch t: Dch t l mt trong nhng bnh nguy him do ly lan nhanh, t l t vong cao. Hin nay, bnh t vn tn pht nhiu tnh nc ta, c bit l vng ng bng sng Cu Long. Ngun ly l phn ngi bnh v ngi lnh mang bnh, nc l yu t lm lan truyn bnh.
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3.3. Chn on vi khun hc

3.3.1. Chn on trc tip Bnh phm l phn hoc cht nn ca bnh nhn, cn ly sm trc khi bnh nhn ung khng sinh. Soi ti v nhum soi: Trn tiu bn soi ti thy vi khun hnh hi cong v di ng, phng php ny quan st tnh di ng ca vi khun t nh sao i ngi, nu quan st knh hin vi nn en. Nhum soi quan st hnh th, tnh cht bt mu ca phy khun t v bch cu trong phn. Nui cy phn lp: + Bnh phm c nui cy vo mi trng pepton kim, sau 6 gi ly vng trn mt mi trng nhum soi v cy chuyn sang cc mi trng phn lp. + ng thi cy bnh phm vo mi trng nh thch kim, hoc TCBS, sau 18 24 gi, nhum soi v xc nh tnh cht sinh ho hc. Phn ng ngng kt: Lm phn ng ngng kt trn lam knh vi khng huyt thanh a gi. Nu ngng kt th tip tc lm phn ng ngng kt vi khng huyt thanh n gi xc nh nhm v typ. K thut khng th hunh quang trc tip: Lm tiu bn t bnh phm hoc t vng mi trng pepton kim, nhum tiu bn bng khng th gn hunh quang ri soi knh hin vi hunh quang. Phng php ny cho kt qu nhanh v tnh c hiu cao. 3.3.2. Chn on gin tip Trn thc t thng khng lm v bnh t c thi k bnh nhanh, khng th c th cha xut hin, kt qu chm. Nhng c th dng iu tra dch t hc.
3.4. Phng bnh v iu tr

Phng bnh: + Phng bnh c hiu: Hin nay c cc loi vaccin dng bng ng ung kch thch p ng min dch ti rut. Vaccin phng t hin nay Vit Nam ang dng gm c O1 v O139 l vaccin bt hot dng huyn dch, a vo c th bng ng ung. + Phng khng c hiu: Nhng bin php phng bnh quan trng l v sinh n ung, pht hin sm v cch ly trit bnh nhn, x l phn v cht nn bnh nhn. Dit rui nhng trung gian truyn bnh. iu tr: + B nc v in gii: C vai tr quan trng nht cu sng bnh nhn, cho bnh nhn ung oresol (ORS) v cc cht lng tng ng vi s nc mt. Truyn tnh mch khi cn thit. + Khng sinh: Thng dng tetraxyclin, cloramphenicol, bactrim.
4. TRC KHUN KHNG CN ACID
4.1. Trc khun lao (Mycobacterium tuberculosis)

Trc khun lao c Robert Koch phn lp c nm 1884 nn cn gi l trc khun Koch (Bacille Koch BK). 4.1.1. c im sinh vt hc 4.1.1.1.Hnh th v tnh cht bt mu Trc khun lao c hnh thng hoc hi cong, mnh, ng ring r hoc thnh tng m. Vi khun khng c lng, khng di ng, khng c v, khng sinh nha bo. Nhum Ziehl Neelsen vi
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khun bt mu .
Hnh 8.11. Trc khun lao (nh di knh hin vi quang hc)
4.1.1.2. Tnh cht nui cy Trc khun lao rt hiu kh. Nhit thch hp l 37OC. Mi trng nui cy i hi phi giu cht dinh dng. Vi khun pht trin rt chm, thng sau 1 2 thng trn mi trng mi c khun lc. Trn mi trng c Loewenstein, vi khun mc sau khong 1 thng, khun lc dng R, kh, x x, mu trng vng ging nh chic sp l. Trn mi trng lng Sauton, trc khun lao mc thnh vng nhn nheo, kh v dnh vo thnh bnh, y c lng cn. 4.1.1.3. Sc khng Trc khun lao c sc khng cao vi cc yu t l ho. Vi khun b tiu dit nhit 70 O 80 C trong 10 pht. Trong m m, vi khun c th sng c mt thng, trong sa c th sng c nhiu tun. Cc thuc st khun nh cresyl, javel, formaldehyd c th tiu dit c vi khun. Vi khun ngy cng khng cc thuc chng lao nh rifampicin, streptomycin, ethabutol, INH,... 4.1.1.4. c t Trc khun lao khng c ni v ngoi c t, hin nay cha xc nh c yu t c lc ca trc khun, nhng c th l tp hp ca nhiu yu t trong yu t si v lp sp vch t bo c ngha quan trng. 4.1.2. Phn loi Gy bnh lao cho ngi gm M. tuberculosis (trc khun lao ngi); M. bovis (trc khun lao b); M. avium (trc khun lao chim). Chng c phn bit vi nhau bi cc tnh cht:
4.1.3. Kh nng gy bnh 4.1.3.1. Gy bnh cho ngi Bnh lao l mt bnh x hi, ly lan d cc nc km pht trin. Bnh lao c thy trn 50% cc bnh nhn AIDS. Trc khun lao xm nhp vo c th ch yu theo ng h hp qua cc
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git nc bt gy lao phi (chim 90% cc th lao), cc m ca ph nang b vi khun xm nhp to ra cc cha vi khun, sau n hch lympho trong vng ri n cc m khc. Trc khun lao c th xm nhp vo c th qua ng tiu ho (thng qua sa b ti) gy lao d dy, rut. T cc c quan b nhim lao u tin, trc khun lao theo ng mu hay ng bch huyt i khp c th gy bnh lao th pht nh lao mng no, lao mng bng, lao xng, khp, hch, thn,... 4.1.3.2. Gy bnh thc nghim Chut lang l sc vt thng c dng nht gy bnh, thc nghim. Ngoi ra th, kh cng cm nhim i vi trc khun lao. 4.1.4. Min dch Sau khi khi bnh lao, ngi bnh c c min dch dch th v min dch t bo nhng khng th khng c vai tr bo v. Phn ng Mantoux l mt test ni b nh gi min dch lao, bn cht l phn ng qu mn mun c nguyn l l khi a mt lng khng nguyn tuberculin tinh ch (t trc khun lao) vo trong da bnh nhn. nhng bnh nhn mc lao hoc nhng ngi nhim lao th sau 72 gi ti ni tim s xut hin mt phn ng d ng c hiu trong da vi biu hin ni qung quanh ch tim, nn cng, ng knh to nh khc nhau tu mc . Nu ng knh t 1cm tr ln l phn ng dng tnh, tc l c th c min dch i vi trc khun lao. ng knh nh hn 1cm l phn ng m tnh, c th cha c hoc cha y min dch vi vi khun lao. C th nhng ngi ang b bnh lao nhng c th suy gim min dch hoc ang b lao nng c th suy kit th phn ng cng m tnh. 4.1.5. Chn on vi khun hc 4.1.5.1. Chn on trc tip Bnh phm: C th l m, phn, nc no tu, nc tiu, tu theo tng th bnh lao. Bnh phm l m c x l vi ho cht lm lng m v dit tp khun. Sau ly tm ly cn nhum soi hoc nui cy. Nhum soi trc tip: Lm tiu bn t bnh phm, nhum Ziehl Neelsen pht hin vi khun khng cn acid AFB (Acid Fast Bacillus). Kt hp vi cc du hiu lm sng v X quang th rt c gi tr chn on. Nhum soi trc tip l phng php c s dng ch yu trong chn on bnh lao phi. Nui cy phn lp: Bnh phm sau khi c x l c nui cy trn mi trng lng Sauton hoc mi trng c Loewenstein, hoc c 2 mi trng, nui cy cho kt qu chnh xc nhng chm, nn chn on thng da vo cc k thut khc. Hin nay, mt s mi trng nui cy nhanh ang c nghin cu s dng vo chn on bnh lao. 4.1.5.2. Gy bnh thc nghim Tim truyn chut lang, hin nay t dng. 4.1.5.3. Phn ng khuch i gen PCR Phn ng cho kt qu nhanh, chnh xc, p dng tt cho cc chn on lao ngoi phi nhng ch thc hin c c s c iu kin. y l k thut c nhy v c hiu cao nhng t. 4.1.6. Phng bnh v iu tr Phng bnh: + Phng c hiu: Vaccin BCG (Bacillus Calmette Guerin) c hai nh bc hc Calmette v Guerin iu ch vaccin bng cch nui cy trc khun lao b nhiu ln trn mi trng c mt b, lm cho trc khun ny mt kh nng gy bnh nhng cn sng v gy c min dch tt. Vaccin ny c dng cho tr s sinh trong chng trnh tim chng m rng, tr ln v ngi ln ch dng khi phn ng Mantoux m tnh. + Cc bin php khc: Nng cao i sng vt cht v tinh thn, ci thin iu kin sng v lm vic. Pht hin sm bnh nhn lao v iu tr trit .
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iu tr: Cc thuc iu tr bnh lao nh INH, rifampicin, pyrazinamide, ethambutol c dng phi hp v iu tr trong thi gian di 6 9 thng.
4.2. Trc khun phong (hi) (Mycobacterium leprae)

Trc khun phong c Hansen, ngi Nauy pht hin nm 1874, nn cn c gi l trc khun Hansen (Bacillus Hansen BH). 4.2.1. c im sinh vt hc 4.2.1.1. Hnh th v tnh cht bt mu Trc khun phong c hnh dng ging trc khun lao nhng mp hn. Trn tiu bn thng thy vi khun ng vi nhau thnh tng m nh b ci. Trc khun phong khng c v, khng c lng, khng di ng, khng sinh nha bo.
Hnh 8.12. Trc khun phong
4.2.1.2. Tnh cht nui cy Hin nay cha nui cy c trc khun phong trn mi trng nhn to. Khi tim truyn vo chut Hamster, trc khun phong sinh sn nhanh ti ch tim, nh vy c th gi chng vi khun nghin cu. 4.2.1.4. c t v enzym Cha hiu bit y v c t v enzym, tuy nhin pht hin c mt s cht gy d ng ca trc khun phong. 4.2.2. Kh nng gy bnh Trc khun phong gy bnh t nhin cho ngi. Vi khun xm nhp vo c th ch yu qua ng da, c th ng qua nim mc. Bnh phong c ly nhng kh ly hn bnh lao, khng di truyn. Thi gian bnh trung bnh 3 5 nm. C trng hp bnh rt di ti vi chc nm. Cc th bnh: Bnh phong c 3 th lm sng: Phong c tnh (phong u): C nhng thng tn ln da, nim mc v cc c quan. Tn thng thn kinh nng, dn n teo c, co rt cc ngn tay, chn, rng cc t xng gy tn ph. Tn thng to thnh u cc ln, nht l vng sn mi khin bnh nhn c b mt d dng. Th ny trong nc mi c nhiu trc khun, kh nng ly mnh nht. Phong c: Th ny cn gi l phong lnh tnh, tn thng thng thy da v nim mc, tn thng thn kinh nh, bnh tin trin chm. Phong bt nh: Cn gi l phong trung gian. Biu hin lm sng trung gian gia hai th trn. C th tr thnh phong c tnh hoc phong c. 4.2.3. Min dch Min dch trong bnh phong l min dch qua trung gian t bo. Dng phn ng Mitsuda nh
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gi v tin lng bnh phong. 4.2.4. Chn on vi khun hc Bnh phm: Ly dch tit hc mi, da vng tn thng,... Nhum soi trc tip: Tiu bn nhum Ziehl Neelsen c th thy trc khun hi ng thnh tng m, nh b ci, bt mu nht. 4.2.5. Phng bnh v iu tr Phng bnh: + Cch ly bnh hi c tnh kp thi. Cc th khc c th iu tr ti nh hoc ti cc tri iu tr tp trung. + Tim vaccin BCG cng c tc dng phng bnh hi. + Ci thin i sng cng ng, v sinh c nhn tt. iu tr: Trc y ngi ta quan nim bnh phong khng cha c. Hin nay c nhiu thuc iu tr khi hon ton bnh phong. Thuc thng c dng iu tr l DDS (Diamino Diphenyl Sulfon).
5. XON KHUN GY BNH

Xon khun gy bnh c gii thiu nm trong b Spirochaetales, gm h Spirochaetaceae v Leptospiraceae. Chng c c im chung sau: Hnh th mnh, xon l xo, mm mi. Nhum Fontana Tribondeau c mu vng nu trn nn vng. Sc khng yu, nhy vi ho cht st trng, khng sinh. Phn loi: B Spirochaetales c 3 ging i din: + Ging Treponema i din l T. pallidum gy bnh giang mai. + Ging Borrelia, i din l B. recurrentis gy st hi quy. + Ging Leptospira gy bnh Leptospirosis.
5.1. Xon khun giang mai (Treponema pallidum)

Nm 1905, Schaudin v Hoffmann tm thy vi khun trong dch tit vt lot ca bnh nhn giang mai. 5.1.1. c im sinh vt hc 5.1.1.1. Hnh th v tnh cht bt mu Xon khun giang mai rt mnh, hnh xon nh l xo, thng c 8 14 vng xon u n. Soi ti trn knh hin vi nn en thy xon khun chuyn ng quay trn. Nhum bng phng php Fontana Tribondeau xon khun c mu vng nu trn nn vng.
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Hnh 8.13. Treponema pallidum (trn tiu bn nhum)
5.1.1.2. Tnh cht nui cy Hin nay cha c mi trng nhn to nui cy xon khun giang mai. Nm 1948, Nelson v Mayer iu ch mi trng c bn c th gi xon khun sng c vi ngy lm cc phn ng huyt thanh c hiu. Phng php gi chng ca Nichols t nm 1911 n nay vn l tim truyn lin tc vo tinh hon th. 5.1.1.3. Sc khng Vi khun b tiu dit bi cc cht st khun thng thng. D cht nhit phng, c bit iu kin kh. 50OC, vi khun cht sau 1 gi. 5.1.2. Kh nng gy bnh 5.1.2.1. Gy bnh cho ngi Giang mai mc phi: Giang mai l mt bnh x hi hin nay rt t gp. Xon khun xm nhp vo c th ch yu qua ng tnh dc. C th ly nhim qua da sy st, nim mc mt, ming nhng rt him. Bnh din bin qua 3 thi k: + Giang mai thi k I: T 10 30 ngy sau nhim khun, xut hin cc vt lot (chancre) b phn sinh dc. Vt lot nng, nn cng, khng nga, khng au, trong c nhiu xon khun. Thi k ny d ly lan. Nu khng c iu tr th sau vi tun vt lot cng lnh khng li so. Vi khun s tip tc vo mu. + Giang mai thi k II: Bt u t 4 8 tun sau khi c vt lot. Tn thng ngoi da l cc dt mu hoa o (gi l o ban). Trong cc dt c t vi khun nhng vn c kh nng ly lan. + Giang mai thi k III: Sau thi gian t vi nm ti vi chc nm, tn thng n su vo t chc to nn cc "gm" (gumma) giang mai da, xng, gan, c bit l tn thng tim mch v h thn kinh. Rt t khi thy vi khun trong cc "gm" giang mai. Giang mai bm sinh: Ph n b bnh giang mai khi c thai t thng th 4 tr, i xon khun c th qua nhau thai vo thai nhi gy sy thai, thai cht lu, qui thai hoc giang mai bm sinh: tr ra c nhng mn phng bn tay, bn chn trong c nhiu xon khun. C th giang mai chm pht sau 5 6 nm, thm ch khi trng thnh mi biu hin bnh. 5.1.3. Chn on vi khun hc 5.1.3.1. Chn on trc tip Thng p dng cho giang mai thi k I, ly bnh phm l cht tit cc vt lot b phn sinh
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dc. Soi ti trn knh hin vi nn en. Nhum Fontana Tribondeau. Nu kt qu (+) r, kt hp vi tin s v triu chng lm sng th c th kt lun c bnh. 5.1.3.2. Chn on gin tip Vic tm khng th trong huyt thanh thng c tin hnh giang mai thi k II hoc III. Cc phn ng dng khng nguyn khng c hiu: S dng khng nguyn l cardiolipin chit xut t tim b nhng c cu trc gn ging cht lipoid ca xon khun giang mai, pht hin khng th reagin trong huyt thanh bnh nhn. Vi khng nguyn cardiolipin c th lm cc phn ng sau: + Phn ng ln bng VDRL(Veneral Disease Research Laboratories). + Phn ng kt hp b th BW (Bodet Wassemann). V khng nguyn khng c hiu nn c th c trng hp (+) gi i vi mt s bnh nh st rt, thn h nhim m hoc ph n c thai trn 7 thng. Do vy, phi lm phn ng 2 ln nhm kim tra kt qu. Phn ng dng khng nguyn c hiu c nhy cao, chnh xc: + Phn ng TPI (Treponema pallidum Immobilisation): Phn ng bt ng xon khun giang mai. + Phn ng FTA (Fluorescence Treponema Antibody): Phn ng min dch hunh quang gin tip. + Phn ng TPHA (Treponema Pallidum Haemagglutination): Phn ng ngng kt hng cu th ng. 5.1.4. Phng bnh v iu tr Phng bnh khng c hiu: + Gii quyt cc t nn x hi. + Gio dc np sng lnh mnh, tnh yu chung thu. + Gio dc tnh dc an ton. + Pht hin sm v iu tr kp thi bnh nhn giang mai. iu tr: penicillin c tc dng iu tr rt tt tiu dit c xon khun.
5.2. Xon khun Leptospira

Bnh do xon khun Leptospira do Weil (ngi c) pht hin nm 1886 nhng n nm 1915 mi tm thy xon khun. 5.2.1. c im sinh vt hc 5.2.1.1. Hnh th v tnh cht bt mu Vi khun hnh si di, rt mnh, 2 u cong nh mc cu. Soi ti trn knh hin vi nn en thy xon khun di ng mnh. Nhum thm bc Fontana Tribondeau, vi khun bt mu vng nu trn nn vng.
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Hnh 8.14. Leptospira trn tiu bn nhum
5.2.1.2. Tnh cht nui cy Leptospira l xon khun duy nht nui cy c trong iu kin hiu kh. Thng nui cy xon khun trong mi trng lng c thm huyt thanh ti ca th. Leptospira pht trin chm, sau 5 10 ngy lm vn nh mi trng nh khi thuc l. 5.2.1.3. Sc khng Leptospira c sc khng yu, b tiu dit 50OC/10 pht, cht nhanh cc mi trng acid, nh sng v cc thuc st khun thng thng d tiu dit c xon khun. Tuy nhin, xon khun chu c lnh, trong nc sng c khong 3 tun. Sng dai dng trong bn ly, nc ng, thch hp nht l nc cng rnh, ng rung, khe sui. 5.2.2. Kh nng gy bnh 5.2.2.1. Gy bnh cho ngi cha mm bnh thng xuyn l cc loi gm nhm v chng o thi vi khun qua nc tiu. ng ly truyn ch yu qua da sy st, vt thng hoc nim mc do tip xc trc tip hoc gin tip vi ngun ly. V d, tip xc trc tip vi gia sc b bnh nh bc s th y, ngi chn nui, cng nhn l m; hay nhim khun gin tip qua nc, t b nhim khun. Bnh Leptospirosis din bin qua 2 thi k: Thi k 1: Thi gian bnh 1 2 tun. Sau , st cao t ngt ko di 3 8 ngy. Triu chng chnh l au c, sung huyt v xut huyt da, nim mc. Thi k ny trong mu c nhiu vi khun.
Thi k 2: Xon khun khu tr v gy tn thng cc c quan ni tng. Biu hin lm sng l hi chng nhim trng nhim c, hi chng gan mt nh vng da, vng mt, nc tiu vng. Ngoi ra, cn cc biu hin tn thng thn, mng no v xut huyt da, nim mc hoc ni tng. Bnh mang tnh cht ngh nghip, thng gp hn nhng ngi lm ngh chn nui, git m
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gia sc, nng dn, cng nhn a cht, lm nghip. Sau khi khi bnh, c th c min dch vng bn. 5.2.2.2. Gy bnh thc nghim Sc vt cm nhim vi xon khun Leptospira l chut lang. 5.2.3. Chn on vi khun hc 5.2.3.1. Chn on trc tip Bnh phm: Thi k 1: Bnh phm l mu lc bnh nhn ang st. Thi k 2: Bnh phm l nc tiu, dch no tu, dch mng bng. Soi ti: Qua knh hin vi nn en thy xon khun di ng. Nhum Fontana Tribondeau. Nui cy bnh phm vo mi trng c bn l Terskikh, EMTH thm 10% huyt thanh th. Tim truyn cho chut lang. 5.2.3.2. Chn on gin tip Phn ng ngng kt tan Martin Pettit. Min dch hunh quang hoc phn ng ELISA. 5.2.4. Phng bnh v iu tr Phng bnh: + Phng c hiu: Dng vaccin cht cho cc i tng tip xc vi mm bnh, c nguy c mc bnh. + Phng khng c hiu: Ct t dy chuyn dch t bng cch dit chut, pht hin v iu tr kp thi cho gia sc b bnh. Trang b phng h lao ng cho ngi tip xc vi ngun ly. iu tr: Dng khng sinh dit xon khun nh penicillin, kt hp vi iu tr triu chng.
6. TRC KHUN UN VN (Clostridium tetani)

Vi khun k kh l nhng vi khun khng sng c khi c mt ca oxy t do, chng s dng c cc hp cht dinh dng nh h thng enzym. C nhiu vi khun k kh khc nhau: cu khun, trc khun, phy khun v xon khun. C loi c nha bo (cc Clostridium), c loi khng c nha bo. Cc vi khun k kh c nha bo c vai tr rt ln trong cc bnh l ca ngi. T chc Y t th gii xp un vn l bnh t ng vt truyn cho ngi v tc nhn gy bnh thng gp phn cc loi gia sc ln. Nm 1889, Kitasato phn lp c vi khun, xc nh c c t v gy bnh thc nghim thnh cng.
6.1. c im sinh hc
6.1.1. Hnh th v tnh cht bt mu Trc khun thng v mnh, khng c v, c lng v di ng, bt mu Gram(+). Khi gp iu kin khng thun li trc khun c kh nng hnh thnh nha bo lm vi khun ging hnh vt.
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Hnh 8.15. Trc khun un vn
6.1.2. Nui cy Trc khun k kh tuyt i, pht trin trn mi trng n gin, nhit 37OC, pH= 7. C th nui cy trc khun trong cc mi trng sau: canh thang glucose, canh thang tht bm hoc gan cc, mi trng thch su. 6.1.3. c im ho sinh Trc khun un vn lm lng gelatin chm, khng phn gii protein, sinh indol, ln men yu cc loi ng: galactose, lactose, saccarose, arabinose, c kh nng gy tan mu. 6.1.4. Kh nng khng Trc khun un vn b tiu dit 56OC sau 30 pht, nhng th nha bo th phi 120OC sau 30 pht mi cht, phenol 5% trong 5 gi, nha bo c th tn ti nhiu nm trong t. 6.1.5. c t c t ca trc khun un vn l mt ngoi c t, bn cht l mt protein c c tnh cao. c t ny gm hai phn: Tetanolysin: Tc dng lm tan hng cu ca th, ngi v nga. c t ny c vai tr rt ph trong gy bnh. Tetanospasmin l c t thn kinh gy nn nhng triu chng c trng ca bnh un vn. y l loi c t c tnh khng nguyn mnh, v vy c th dng sn xut vaccin.
6.2. Min dch

S hnh thnh min dch hot ng v bn vng do tim vaccin gii c t un vn (anatoxin). Ngi m mang thai t thng th 6 c tim vaccin un vn, khi p ng min dch c hnh thnh, khng th s c truyn cho thai nhi qua nhau thai gip phng bnh un vn rn.
6.3. Kh nng gy bnh

6.3.1. Gy bnh cho ng vt Trc khun un vn thng gy bnh cho cc loi ng vt c v nh: tru, b, nga, ch, mo v mt s ng vt nh nh th, chut, 6.3.2. Gy bnh cho ngi: Bnh un vn Vi khun hoc nha bo xm nhp vo c th qua nhng vt thng ngoi da su v kn. Ti , vi khun nhn ln v tit c t. Bnh un vn ngi l hin tng nhim c t. Thi gian nung bnh t 5 10 ngy, i khi lu hn. Triu chng khi u l au v cng c
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ni b thng nhng thng b b qua. Triu chng r rt sm nht l cng hm do co cng c nhai, sau l co cng c mt lm bnh nhn kh h mm v nt mt thay i. Tip n l co cng cc c gy, lng, ngc, bng v cc c chi lm cho lng v c ngi un cong ln, v vy gi l bnh un vn. Giai on cui ca bnh, s co tht lan rng ra cc c h hp lm bnh nhn nut v th kh khn, cc chc nng tun hon, h hp b ri lon. Nhng cn co git lin tip xut hin lm bnh nhn v cng au n v thng t vong do suy h hp. c t thn kinh thng lm thn nhit tng cao, c khi n 41 OC, mch nhanh t 150 n 180 ln/pht, huyt p gim,.... Dch t: a s cc ng vt nh tru, b, nga, cu, ln, ngay c ch, mo, chut c th mang vi khun un vn k sinh trong rut. Nu iu kin v sinh thp km th vi khun c kh nng ly lan ra mi trng ngoi, ri t xm nhp vo c th ngi qua cc vt thng ngoi da.
6.4. Chn on
chn on bnh un vn ch yu da vo du hiu lm sng. Chn on vi sinh t lm, nu c th ch p dng nhng c s c y iu kin nui cy vi khun k kh.

Phng bnh: + V sinh mi trng, x l phn gia sc. + Trng hp vt thng c kh nng nhim un vn, cn x l ngoi khoa nh: ra sch, ct lc, rch rng,... v tim khng huyt thanh un vn SAT (serum anti tetani). + Tim vaccin un vn cho tr em v ph n mang thai. iu tr: + Ra sch vt thng. + Trung ho c t bng huyt thanh khng un vn SAT. + Chng co git bng cc thuc an thn, dn c, trnh cc kch thch thn kinh nh nh sng, ting ng mnh,... + Khng sinh dit khun. + Ch chm sc c bit.
7. TRC KHUN THAN (Bacillus anthracis)

Bacillus anthracis thuc h Bacillaceae chi Bacillus, chi ny gm nhiu loi trong ch c Bacillus anthracis gy bnh cho ngi. Bacillus anthracis c Davaine v Rayer pht hin nm 1850.
7.1.1. Hnh th v tnh cht bt mu Trc khun than c hnh gy, 2 u vung, ng ring r hoc xp vi nhau thnh chui, nhum bt mu Gram (+), nha bo hnh trng hoc hnh tr, thng gia v khng lm bin dng thn vi khun.
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Hnh 8.16. Trc khun than
7.1.2. Tnh cht nui cy Trc khun than hiu kh, pht trin d dng trn cc mi trng nui cy thng thng, nhit 35OC v pH 7 7,4. 7.1.3. Tnh cht sinh ho hc Ln men khng sinh hi ng glucose, maltose, levulose, saccarose. Khng ln men ng lactose, galactose, arabinose. C kh nng ly gii protein, lm lng gelatin. 7.1.4. c t c t ca trc khun than gm c ni c t v ngoi c t. 7.1.5. Cu trc khng nguyn Khng nguyn v l polypeptid. Khng nguyn thn l polyozid. Khng nguyn phc hp ho tan c kh nng min dch khi bo ch thnh vaccin. 7.1.6. Sc khng Trc khun than d b cht 58OC/1 gi, nhy cm vi cc cht st khun thng thng. Nha bo b cht 100OC sau 10 pht hoc cc cht giu oxy (H2O2, thuc tm). Nha bo tn ti 20 30 nm trong t.
7.2. Kh nng gy bnh
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7.2.1. Gy bnh cho ng vt Trc khun than gy bnh cp tnh ng vt n c, c th gy bnh cho mt s ng vt khc. Thng gp th nhim khun huyt gy t vong. Sau khi sc vt cht, d c chn su nhng nha bo c th ln mt t do nhiu nguyn nhn nn nha bo c trong cy c v sc vt n c s d mc bnh. 7.2.2. Gy bnh cho ngi Ngi b mc bnh than do tip xc vi da hay tht, lng ng vt b bnh. Bnh thng gp 3 th: Th da: Vi khun xm nhp qua da do tip xc gy tn thng ti ch. Da xut hin nt phng, sau , gia c mu en ri hoi t. Bnh hay gp cng nhn l m v thuc da. Th ny chim ti 90%. Th phi: Ngi mc bnh do ht phi nha bo vo ng h hp gy vim phi nng, vim thn, nhim c, c th nhim khun huyt v t vong. Th ny chim khong 5%. Th rut: do bnh nhn nhim vi khun bng ng tiu ho, gy xut huyt v hoi t hch lympho rut v hch mc treo, c th tn thng hch lympho hu, hng. Th ny rt nng nhng t khi gp. cc th bnh trn, a s bnh nhn nhim nha bo, sau khi vo c th, nha bo tr thnh th hot ng gy ph keo t chc v sung huyt cc m. Trc khun than ti cc hch lympho, lch ri n mu, t mu n cc t chc c quan sau khi nhn ln rt nhanh mu. Do to thnh nha bo c kh nng khng cao nn vi khun than c dng lm v kh sinh hc. Vn ny xy ra nc M nm 2001. 7.2.3. Gy bnh thc nghim Sc vt cm nhim l chut nht, chut lang. Chut cht sau khi a mt liu nh vi khun vo c th.
7.3. Chn on vi sinh hc

7.3.1. Chn on trc tip Bnh phm: Tu thuc vo th bnh m bnh phm c th l cc nt m da, m, mu. Nhum soi: C gi tr nh hng, nhum Gram tm vi khun. Nui cy: + Bnh phm l m, m, cht tit nui cy vo mi trng thng thng hoc thch mu, 35 O 37 C, sau 24 48 gi, quan st khun lc v xc nh cc tnh cht sinh ho hc. + Bnh phm l mu: Cy mu tt c bnh nhn mc bnh than, phi theo di t nht 1 tun, nu c vi khun th xc nh tnh cht sinh ho hc. 7.3.2. Gy bnh thc nghim Tim trc tip bnh phm vo di da hoc phc mc ca chut lang, chut cht sau 24 72 gi do nhim khun huyt. 7.3.3. Chn on gin tip Tm khng th trong huyt thanh bnh nhn.

Phng bnh: + Pht hin sm ng vt mc bnh, cch ly v iu tr kp thi, nu ng vt cht, cn chn su v x l cng ho cht nh vi bt, chn xa cc ngun nc, bi c.
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+ Trang b bo h lao ng cho cng nhn thuc da, ngi git m ng vt. + m bo v sinh mi trng c s st sinh, thuc da. Dng vaccin cho cc i tng c nguy c. iu tr: Penicillin l thuc c tc dng tt vi trc khun than.
8. VI KHUN HAEMOPHILUS INFLUENZAE
8.1.1. Hnh th v tnh cht bt mu H. influenzae l vi khun a hnh thi, nhum bt mu Gram(). Khng c lng, khng di ng, c th c v, khng sinh nha bo.
Hnh 8.18. Haemophilus influenzae (nh di knh hin vi quang hc)
8.1.2. Tnh cht nui cy Vi khun hiu kh, ch pht trin trn mi trng c 2 yu t X v V. Yu t X bao gm cc cht mu cha st nh Hemin, Hematin. Yu t V l NAD (nicotinamide dinucleotide) nm trong t bo mu. Nhit thch hp 37OC, kh trng 5%CO2. 8.1.3. Tnh cht sinh ho hc Ln men ng glucose. Catalase (+). 8.1.4. Cu trc khng nguyn H. influenzae c khng nguyn v c bn cht l polysaccharid. Da vo khng nguyn v chia 6 typ huyt thanh: a, b, c, d, e, f. Typ b thng gp gy bnh. V c vai tr quan trng gip vi khun chng li hin tng thc bo. V ca typ b c dng sn xut vaccin. 8.1.5. Sc khng H. influenzae c sc khng rt yu. 8.2. Kh nng gy bnh H. influenzae v ph cu l hai vi khun hng u gy nhim khun ng h hp tr em di 5 tui. H. influenzae k sinh bt buc ng h hp ca ngi, gp nhiu tr em, khong 75% tr c mang vi khun ny mi hng nhng khng gy bnh. Bnh do H.influenzae thng gy bnh th pht sau bnh cm, si, gp nhiu nht tr 2 thng 5 tui. Vi khun c th gy nhim khun ng h hp trn, vim hng, vim mi, vim xoang, vim tai gia, c bit, gy vim phi v vim mng phi. Vi khun c th xm nhp vo mu ri n mng no
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gy vim mng no. Vim mng no tr nh do H. influenzae thng rt nng, cn c chn on sm v iu tr kp thi.

8.3.1. Chn on trc tip Bnh phm: Dch mi hng, dch no tu, mu,... Nhum soi quan st hnh th. Nui cy bnh phm vo mi trng thch hp. Sau 18 24 gi nhum soi khun lc v xc nh cc tnh cht sinh ho hc. Tm khng nguyn v typ b trong bnh phm bng k thut min dch. 8.3.2. K thut PCR pht hin on ADN c hiu

Phng bnh: + Phng c hiu vaccin c tinh ch t v polysaccharid ca H. influenzae typ b. + Phng khng c hiu: Bnh nhn vim mng no do H. influenzae phi c cch ly, dng khng sinh d phng cho nhng ngi c tip xc. iu tr: Hin nay c nhiu chng H. influenzae khng li cc thuc trn nn tt nht iu tr theo khng sinh . Loi khng sinh cn tc dng tt vi H. influenzae l cephalosposin nh cefotaxin, ceftriaxon.
9. TRC KHUN BCH HU (Corynebacterium diphteriae)

Nm 1884, Loffler phn lp c trc khun bch hu v xc nh vai tr gy bnh, do , trc khun bch hu cn mang tn l vi khun KlebsLoffler.
9.1.1. Hnh th v tnh cht bt mu Trc khun hnh thng, c 1 hoc 2 u phnh to nh hnh try, khng v, khng sinh nha bo. Nhum Gram bt mu Gram(+), nhum Albert vi khun bt mu xanh.
Hnh 8.19. Trc khun bch hu (nh di knh hin vi in t)
9.1.2. Tnh cht nui cy Trc khun pht trin trn mi trng giu cht dinh dng, nhit thch hp l 37OC. Mi trng nui cy thng dng l: mi trng huyt thanh ng (Loffler), mi trng trng, mi trng Schroer (thch mu c mui kali telurit).
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9.1.3. Tnh cht sinh ho hc Enzym urease (), y l tnh cht quan trng phn bit vi trc khun gi bch hu. Catalase (+), indol (). Ln men khng sinh hi ng glucose, maltose. Khng ln men ng lactose. Da vo tnh cht sinh ho hc phn bit trc khun bch hu v trc khun gi bch hu c hng:
Gia cc trc khun bch hu c s khc nhau v c im sinh hc, trn c s ny chia thnh 3 typ sinh hc l: Gravis, Mitis v Intermedius, vi cc c im sau:
Ba typ sinh hc trn c s khc nhau v kh nng gy bnh: Gravis thng gy dch bch hu ln, cn Mitis gy dch tn pht, nhng tn ti dai dng. Dch bch hu nc ta thng do Mitis. 9.1.4. Cu trc khng nguyn C 2 loi: Khng nguyn thn O v khng nguyn b mt K. 9.1.5. c t Ngoi c t, bn cht l glycoprotein. Ngoi c t bch hu c x l bng formalin 0,5% 37OC th khng cn kh nng gy bnh nhng cn tnh khng nguyn v c dng iu ch vaccin phng bnh bch hu rt hiu qu. 9.1.6. Sc khng Tng i cao, vi khun c th sng c 2 3 tun trong dng dng c, 5 tun trong bi. B cht 58OC sau 10 pht, 100OC sau 1 pht. B cht di nh sng mt tri trong vi gi.
9.2. Kh nng gy bnh

Trc khun bch hu xm nhp vo c th bng ng h hp v c tr phn trn ng h hp, thng gp nht l vng hu hng. Trc khun pht trin tit ra ngoi c t gy hoi t lp biu m, kch thch tng xut tit to ra mng gi. Mng gi trng xm, dai, kh bc tch, khi bc hay chy mu. Mng gi c th lan xung thanh qun, kh qun gy bt tc ng h hp. Ngoi c t bch hu thm vo mu gy nhim c ton thn. Cc c quan b tn thng nng do ngoi c t bch hu l tim (bnh nhn thng cht do bin chng tim), thn kinh ngoi bin (c th gp bin chng lit), tuyn thng thn v gan. Bnh bch hu thng gp tr em, sau khi khi bnh, c th thu c min dch. y l mt bnh dch nguy him tr em.
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Bnh phm: Mng gi bch hu, dch ngoy hng. Nhum soi: Bnh phm thng c nhum bng phng php Gram v Albert hoc xanh methylen. Nui cy phn lp: Bnh phm c nui cy vo mi trng phn lp thch hp nh mi trng trng, mi trng huyt thanh ng, mi trng Schroer. Nhum soi khun lc, nu l trc khun Gram(+) th kim tra cc tnh cht ca trc khun bch hu. Xc nh c t bch hu: khng nh trc khun bch hu c c lc, cn xc nh c t ca chng, c th dng 2 phn ng sau: + Phn ng trung ho trn chut lang. + Phn ng Elek.

Phng bnh: + Phng c hiu: Vaccin l gii c t bch hu. + Phng khng c hiu: Pht hin sm v cch ly trit bnh nhn, dp tt kp thi khi c dch xy ra. iu tr: + Dng khng c t bch hu SAD (serum anti diphterique). + Dng khng sinh dit khun: Tt nht l penicillin, erythromycin. + Ch iu dng thch hp phng bin chng tim mch.
10. TRC KHUN HO G (Bordetela pertussis)

Trc khun ho g thuc chi Bordetela h Alcaligenaceae do J. Bordet v O. Gengou phn lp t mt bnh nhi b ho g nm 1906.
10.1.1. Hnh th v tnh cht bt mu B. pertussis hnh cu trc khun nh, bt mu Gram(), vi khun khng c lng m c cc si ngng kt hng cu (filamentous hemagglutinin FHA). FHA gip vi khun bm vo t bo c lng chuyn ca biu m h hp.
Hnh 8.20. Trc khun ho g
10.1.2. Tnh cht nui cy
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B. pertussis hiu kh tuyt i, nhit thch hp l 37OC, rt kh nui cy trn cc mi trng thng thng. Mc chm trn mi trng BordetGengou. 10.1.3. Tnh cht sinh ho hc B. pertussis chuyn ho ng theo li h hp, khng bao gi ln men. 10.1.4. Sc khng khng yu vi cc yu t ngoi cnh, cht nhanh khi ra khi c th. 10.1.5. c lc c t ho g (pertussis toxinPT) l ngoi c t c bn cht l protein, gy ra cc triu chng lm sng ca bnh. c t t bo kh qun gy tn thng cc t bo c lng chuyn biu m ng h hp. 10.1.6. Khng nguyn FHA v PT l nhng khng nguyn mnh v quan trng nht, ngoi ra vi khun cn c cc yu t khng nguyn chu nhit.
10.2. Kh nng gy bnh

Gy bnh ho g l bnh nhim trng nhim c h hp cp tnh nng v rt d ly lan tr em. Ngi l vt ch duy nht ca B. pertussis, vi khun k sinh trn nim mc h hp, bm vo cc t bo c lng chuyn bng si ngng kt hng cu v tit cc c t lm tn thng nim mc dn n nhng cn ho d di. Nim mc h hp b tn thng thng dn n bi nhim vi khun gy vim phi lm tnh trng bnh cng trm trng. Trong trng hp ho g nng, c th gy h ng huyt v tn thng no do thiu oxy, gy bin chng, nh hng n tr tu ca tr. Tr s sinh khng mc ho g v cn khng th ca m. Nhng nu khng c tim chng th la tui 1 5 d mc bnh nht do lc ny min dch th ng ht trong khi min dch ch ng cha c thit lp. Sau mc bnh c min dch bn vng lu di.

10.3.1. Chn on trc tip Bnh phm l dch mi hng. Nhum soi theo k thut min dch hunh quang trc tip. Nui cy vo mi trng thch hp xc nh tnh cht sinh hc c trng. K thut PCR tm on ADN c trng. 10.3.2. Chn on gin tip Tm khng th khng PT v FHA c hiu trong huyt thanh.

Phng bnh: + Pht hin sm, cch ly bnh nhn. + Tim vaccin cho tr em trong chng trnh tim chng m rng. iu tr: + Khng sinh chn l erythromycin. + Ch chm sc v dinh dng thch hp.
11. TRC KHUN M XANH (Pseudomonas aeruginosa)

Trc khun m xanh thuc h Pseudomonaceae chi Pseudomonas do Schroeter m t nm 1872. Nm 1900, Migula chuyn sang chi Pseudomonas.
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11.1.1. Hnh th v tnh cht bt mu Trc khun thng hoc hi cong, hai u trn. C mt lng mt u, di ng, t khi c v, khng sinh nha bo. Nhum bt mu Gram().
Hnh 8.21. Trc khun m xanh (nh di knh hin vi in t)
11.1.2. Tnh cht nui cy Trc khun m xanh mc d dng trn cc mi trng nui cy thng thng, hiu kh. Nhit thch hp 37OC, pH thch hp 7,2 7,5. Trn mi trng c c th gp 2 loi khun lc: 1 loi to, nhn, dt, trung tm li ln, c xu hng mc lan; 1 loi x x, b khng u, i khi c loi khun lc nhy. Trong mi trng lng vi khun mc vng trn, mi trng c. 11.1.3. Sc t Tnh cht c trng ca trc khun m xanh l sinh sc t v cht thm. C 2 loi sc t chnh: Pyocyanin: C mu xanh l cy. a s trc khun m xanh sinh sc t ny. Sc t ny lm cho m vt thng do trc khun c mu xanh. Pyoverdin l loi sc t hunh quang. Sc t ny khng bn vng, d b mt i trong iu kin nui cy khng tt. Sc t ca trc khun m xanh di nh hng ca cc nhn t ho hc c th thay i thnh sc t nu, en. Cht thm do trc khun m xanh sinh ra l kimetylamin. 11.1.4. Tnh cht sinh ho hc Khng ln men ng lactose. Oxydase (+), catalase (+). Indol (); H2S (). 11.1.5. Cu to khng nguyn Khng nguyn lng H. Khng nguyn thn O. 11.1.6. Sc khng Trc khun m xanh b tiu dit nhit 100OC v cc thuc st khun thng thng.
11.2. Kh nng gy bnh
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Trc khun m xanh thng sng trong t, nc hoc trn da, nim mc ngi v ng vt. L loi vi khun gy bnh c iu kin, khi c th b suy gim min dch, b mc bnh c tnh hoc mn tnh. Trc khun m xanh c nhiu ni, nh trong nhiu dng c my mc trong bnh vin nh ng thng, my h hp nhn to. Chng xm nhp vo c th qua da (nht l sau khi b bng) hoc qua vt thng, do phu thut. Ti ch vi khun gy vim c m, in hnh l m c mu xanh. Nu c th gim sc khng hoc do bnh ton thn, vi khun xm nhp v gy vim cc c quan nh vim bng quang, vim tai gia, vim mng no, vim mng bng,... C th vi khun vo mu gy nhim khun huyt, vim ni tm mc. Ngoi ra, ngy nay trc khun m xanh c coi l tc nhn gy nhim khun bnh vin mc phi nhng bnh nhn nm vin lu ngy. Nhim khun do trc khun m xanh ngy cng tr nn trm trng do s khng khng sinh rt mnh ca vi khun.

Bnh phm: M, dch mng phi, dch mng bng, dch no tu, nc tiu, mu tu theo tng th bnh. Nhum Gram. Nui cy trn mi trng thch hp. Sau 18 24 gi nhn xt khun lc, chn khun lc mu xanh v lm xanh mi trng nhum soi, ri xc nh tnh cht sinh ho hc.

Phng bnh: Ngn nga nhim trc khun m xanh bng cc bin php v sinh, kh trng nht l cc bnh vin v c s y t. iu tr: Trc khun m xanh khng li rt nhiu loi khng sinh. Hin nay, loi khng sinh cn tc dng l amikacin hay cc cephalosporin th h 3. Tt nht cn iu tr theo khng sinh .
12. HELICOBACTER PYLORI

Nm 1979, Warren (Australia) a ra nhn xt, nhng tiu bn gii phu bnh l ca bnh nhn vim lot d dy, t trng c mt loi vi khun hnh xon nm lp nim mc. Nm 1981, Marshall (Australia) bt u phn lp vi khun v chnh thc cng b vi khun ny vo nm 1983. Nm 1989, Goodwin nghin cu v cu trc t bo v ngh t tn l H. pylori.
12.1.1. Hnh th v tnh cht bt mu H. pylori c hnh xon hoc hi cong. Di ng mnh do c chm lng u, khng c v, khng sinh nha bo, bt mu Gram(). 12.1.2. Tnh cht nui cy Nui cy H.pylori rt kh. Mi trng nui cy phi giu dinh dng v mt s yu t c bit, nhit thch hp 37OC v kh trng 7% CO2. Trn mi trng nui cy sau 48 72 gi, khun lc trong hoc xm nht, i khi tan mu ng knh khong 1mm.
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Hnh 8.22. Helicobacter pylori
12.1.3. Tnh cht sinh ho hc Urease(+). Catalase (+), oxydase (+). Nitrat (). 12.1.4. Sc khng Qua cc nghin cu cho thy, thc cht H. pylori khng phi l vi khun a acid v chng b tiu dit rt nhanh mi trng c pH l 3,1 3,5 trong khi chng tn ti lu d dy c pH l 2,5 3,0, v H. pylori c kh nng tit urease rt mnh, phn gii ure trong d dy to thnh amoniac bao quanh vi khun, lm cho vi khun chu c mi trng acid. 12.1.5. Khng nguyn Khng nguyn lng H: Bn cht l protein. Khng nguyn thn O: Bn cht l lipopolysaccharid chu nhit, khng nguyn ny c tnh c vi t bo tc ch. Khng nguyn l cc enzym: Mt s khng nguyn l cc men c vai tr quan trng lin quan n kh nng gy bnh nh urease, catalase, hismatase, adhezin gip vi khun bm vo t bo nim mc. 12.1.6. c t: C 2 loi c t gy lot t trng. c t gy tng tit dch v.
12.2. Phn loi

Hin nay tm thy 15 loi trong ging Helicobacter, trong c 4 loi k sinh ngi nhng ch c H. pylori gy bnh.
12.3. Kh nng gy bnh

c tnh 5 10% dn s th gii b bnh au d dy, t l ny Vit Nam l 7%. im quan trng nht y hc khm ph ra l c n 70% s dn c nguy c mc bnh v nguyn nhn c xc nh l do H. pylori. H. pylori gy vim lot d dy, t trng do tit ra cc enzym lm suy thoi mng nhy bo v dn n vim v lot. H. pylori cn tit ra c t gy c v ph hu t bo v c t gy tng tit dch v. T chc y t th gii xc nh H. pylori cng l mt trong nhng tc nhn gy ung th d dy. Nguy c ung th d dy ngi nhim H. pylori s tng t 6 10 ln so vi ngi khng
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nhim. H. pylori c mt trong khong 65 70% trng hp vim d dy, 70 80% ung th d dy, hn 90% lot d dy hoc t trng. S ly truyn bnh qua ng tiu ho; rui l trung gian truyn bnh quan trng.

12.4.1. Chn on trc tip Bnh phm: Mnh sinh thit vng vim, lot ca d dy, t trng. Nhum soi: Nhum Gram. Nui cy: Nghin bnh phm ri nui cy vo mi trng thch hp, xc nh tnh cht sinh ho hc ca vi khun. K thut khuch i gen PCR: K thut ny cho php pht hin c on gen c hiu ca H. pylori c mnh sinh thit d dy, dch d dy. 12.4.2. Chn on gin tip Test urease trong bnh phm: Da vo kh nng sinh urease rt mnh ca H. pylori. Test urease trong hi th: Pht hin s c mt ca men urease trong trng hp vim nim mc d dy do nhim H. pylori. Tm khng th trong huyt thanh bnh nhn. Phng php ny c gi tr cao trong chn on dch t hc.

Phng bnh: Hin nay cha c vaccin phng bnh. y l bnh ph thuc nhiu vo iu kin kinh t x hi. Ch yu l nng cao i sng, v sinh mi trng, v sinh n ung. iu tr: + Dng khng sinh dit khun: amoxicillin, clarithromycine. + iu tr vim lot v tng tit dch v. + Ch n trnh cc cht kch thch nh chua, cay,....
CU HI LNG GI
1. Trnh by c im sinh vt hc ca t cu vng. 2. Nu 4 loi enzym ng dng trong chn on VSV t cu vng. 3. Nu 4 th bnh do t cu vng gy ra 4. Nu cc bc trong chn on VSV t cu vng. 5. Trnh by c im sinh vt hc ca lin cu. 6. Tn enzym c tc dng lm tan t huyt ca lin cu. 7. Trn mi trng thch mu, lin cu c nhng hnh thc tan mu no? 8. Nu cc bc trong chn on trc tip lin cu nhm A. 9. Nu nguyn l ca phng php chn on gin tip lin cu nhm A. 10. Trnh by c im sinh hc v cc yu t c lc ca ph cu. 11. Nu tn khng nguyn quan trng nh typ ph cu. 12. Nu cc bc trong chn on trc tip ph cu. 13. Trnh by c im sinh hc ca no m cu. 14. Tn khng nguyn quan trng nh nhm no m cu. 15. Trnh by c im sinh hc v kh nng gy bnh ca lu cu.
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16. Phng php chn on trc tip lu cu. 17. Tnh cht ho sinh quan trng phn bit lu cu v no m cu. 18. Trnh by c im sinh hc ca Haemophilus influenza. 19. Nu phng php chn on trc tip Haemophilus influenza. 20. Trnh by c im sinh hc ca Salmonella. 21. K tn 4 loi Salmonella gy bnh thng hn. 22. Trnh by 2 th bnh lm sng do Salmonella. 23. Nu cc thi im ly bnh phm chn on bnh thng hn. 24. Nu phng php chn on vi sinh bnh thng hn. 25. Trnh by c im sinh hc ca Shigella. 26. K tn 4 nhm vi khun Shigella. 27. Nu cc bc trong chn on vi sinh trc khun Shigella. 28. Trnh by c im sinh hc ca E. coli. 29. Nu tn 5 nhm E. coli gy tiu chy. 30. Nu phng php chn on trc tip E. coli. 31. Trnh by c im sinh hc ca phy khun t. 32. Nu cu to khng nguyn v phn loi phy khun t. 33. Nu tnh cht sinh hc quan trng phn bit V. c in v V. Eltor. 34. Nu phng php chn on trc tip trc khun t. 35. Trnh by c im sinh hc ca trc khun lao. 36. Nu phng php chn on trc tip trc khun lao. 37. Trnh by v phng php phng v nguyn tc iu tr bnh lao. 38. Trnh by c im sinh vt hc v kh nng gy bnh ca trc khun phong. 39. Trnh by c im sinh vt hc v kh nng gy bnh ca xon khun giang mai. 40. Trnh by c im sinh vt hc v kh nng gy bnh ca xon khun Leptospira. 41. Trnh by c im sinh vt hc v kh nng gy bnh ca trc khun un vn. 42. Trnh by c im sinh vt hc v kh nng gy bnh ca trc khun bch hu. 43. Trnh by c im sinh vt hc v kh nng gy bnh ca trc khun m xanh. 44. Trnh by c im sinh vt hc v kh nng gy bnh ca vi khun H. pylori.
Chng 9
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VI NM GY BNH
1. M U
Trong nhng nm gn y trn th gii, cc bnh do vi nm gy ra xut hin ngy mt tng. Theo nhiu chuyn gia trong lnh vc lm sng nh gi th mi e do ca vi nm i vi sc khe con ngi ni ring v ng vt ni chung ch ng sau virus v vi khun. S pht trin cc bnh do vi nm gy ra trong vi thp nin tr li y l do nhiu nguyn nhn. Trong mt nguyn nhn quan trng khng th khng ni n, l s bng n ca i dch AIDS. S ly nhim virus HIV lm tn thng h thng min dch v l c hi cho cc bnh nm xut hin. Tip n l cc nguyn nhn nh vic s dng khng sinh, cc cht corticoid mt cch tu tin cng l yu t quan trng lm gia tng cc bnh do nm. Bn cnh , s ngi b bnh tiu ng ngy mt tng (c a ca ngi mc bnh tiu ng d b mc mt s bnh nm), cc ca ghp c quan c bit l ghp thn cng khng ngng tng ln l nhng c hi cho nm ly nhim v gy bnh. Trong khi , vic u t cho nghin cu sn xut cc thuc khng sinh khng nm cha c ch c v thi gian ln kinh ph. Ngoi nhng yu t trn, iu kin kh hu nng m, iu kin sng, iu kin v sinh phng bnh ni chung cn thp nh nc ta cng l nhng yu t thun li cho s pht trin ca cc bnh do vi nm gy ra.
2. C IM V PHN LOI VI NM
2.1. c im ca vi nm
Vi nm thuc nhm sinh vt nhn tht (Eucaryota), c th nm khng c mch, sinh sn bng bo t v thng pht tn vo mi trng nh gi. Bo t ca nm gm 2 loi: bo t hu tnh v bo t v tnh. Cc loi bo t ny c sinh ra tu thuc vo loi v cc iu kin ngoi cnh. Nm ging thc vt u c s thay i th h. Nm khng c kh nng di ng, ngoi tr mt s rt t thuc B Nm roi (Chytridiales) c pha di ng. Th sinh dng ca nm c th l n bo (nm men) hoc dang si (nm mc). Mt s loi nm c dng lng hnh (tn ti c 2 dng trn cc iu kin mi trng khc nhau). Thnh t bo ca nm c cu trc tng t thnh t bo thc vt nhng khc nhau v thnh phn ho hc: thnh t bo nm cu to ch yu l cht chitin, thnh t bo thc vt cu to ch yu l cellulose. Nm khng phi l cc sinh vt t dng m l cc sinh vt d dng. Cng l sinh vt d dng nhng nm khc vi ng vt ch, ng vt nut thc n vo d dy ri mi tiu ho cn nm th ngc li, tit ra cc enzym ngoi bo phn gii cc cht hu c phc tp thnh cc cht n gin ri mi hp thu vo c th. Nm tch tr nng lng di dng glycogen nh ng vt v khc vi thc vt tch tr nng lng di dng tinh bt. Mng t bo nm c mt sterol duy nht, l
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ergosterol, cht thay th cho cholesterol c mng t bo ng vt c v. Hu ht nm c nhn t bo rt nh v ADN t trng lp. Qu trnh nguyn phn ni chung c kt thc m khng c s phn hu mng nhn. Tu thuc vo loi v iu kin ngoi cnh, nm c th c cc trng thi dinh dng: hoi sinh, k sinh v cng sinh. 2.1.1. Hnh dng v cu trc Cc loi nm gy bnh c 2 dng: dng si gi l nm mc, dng n bo gi l nm men. Nm mc pht trin thnh cc ng dng si, nh b, thng khng nhn thy bng mt thng v phn nhnh. Cc on si ny c gi l hyphae v tp trung thnh h si (mycelium). H si l ci m chng ta nhn thy khi chng ta quan st lp mu trng trn cc loi qu b mc. Cc si (hyphae) hoc c vch ngn, hoc dng cng bo (si a nhn nhng khng c cc vch ngn ring tng t bo) (hnh 9.1). y l cc c im hnh thi ca nm c s dng trong chn on xc nh nm cc labo xt nghim. Trn thch, cc si pht trin lan ra t im cy bng vic pht trin cc chp si v sau phn thnh cc nhnh. Dng nm men l cc t bo n, hnh trng hoc hnh cu vi thnh t bo cng v phc hp t bo cng tng t nh dng si. Hu ht nm men phn chia theo kiu ny chi (hnh 9.2), mt s t loi khc phn chia theo kiu phn i (fission) ging vi khun. Trn mi trng thch chng hnh thnh cc khun lc tng t cc khun lc ca vi khun nhng thng ln hn ng k. Mt s to ra cc v (capsule) c cu to bi polysaccharid, mt c tnh quan trng ca loi Cryptococcus neoformans, mt tc nhn gy bnh vim mng no rt thng gp bnh nhn AIDS.
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Hnh 9.2. Sinh sn ny chi ca nm men
2.1.2. Tnh lng hnh v s pht trin Nhiu loi nm gy bnh c kh nng tn ti hai dng hnh thi khc nhau, l kh nng tn ti hoc dng nm si hoc dng nm men tu thuc vo iu kin mi trng pht trin. V d, cc tc nhn ca penicilliosis v histoplasmosis ln lt l Penicillium marneffei v Histoplasma capsulatum pht trin nh mt nm men mt s iu kin v pht trin thnh dng si mt s iu kin khc (hnh 9.3).
Hnh 9.3. Hin tng lng hnh ca Penicillium marneffei
Hin tng ny c gi l tnh lng hnh (dimorphism). iu kin phng th nghim, s chuyn i gia hai pha ny c th c to ra mt cch thun nghch bi s thay i nhit . Pha nm men in hnh hn nhit c th ngi. S chuyn i nm men nm si hoc nm si
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nm men thng xuyn xut hin khi mt loi nm sng t do ngoi mi trng tr thnh mt sinh vt k sinh. Hu ht cc loi nm gy ra cc bnh ton thn nh cc tc nhn ca histoplasmosis v blastomycosis, s c dng nm men khi k sinh ngi, v c dng si khi pht trin ngoi mi trng (ch yu l trong t). Trong mt s t trng hp khng bnh thng nh Candida quy lut ny b o ngc, dng si (ng hn l gi si) thng tm thy trong cc m ca vt ch (hnh 9.4). Khng phi tt c cc loi nm gy bnh u tn ti di dng lng hnh v tri qua s bin i hnh thi khi chng xm nhim vo c th: chi Aspergillus l mt trong s nm mc ph bin nht ngoi mi trng v lun tn ti dng nm si. Tri li loi Cryptococcus neoformans th lun tn ti dng nm men. Mt s loi nm men nht nh, c bit l cc loi ca chi Candida xut hin mt dng ny chi bin i, trong , cc t bo con mi c to thnh do ny chi vn dnh vo cc t bo m v tr thnh chui di ging nh si. S kt hp ny c gi l cc si gi hay ging si (pseudohyphae), hoc to thnh l h si gi (pseudomycelium). Hu ht cc vn v sinh hc phn t v di truyn ca cc loi nm c nghin cu ng dng, c bit l nm men bia hoc nm men bnh m (bakers or brewers yeast) Saccharomyces cerevisiae v tm hp hn l cc loi nm si nh Aspergillus nidulans v Neurospora crassa. t c bit hn l cc loi nm gy bnh mc du nhiu cng trnh nghin cu ang c tin hnh v cc kha cnh nh kh nng gy bnh, mc nguy him ca bnh do chng gy ra v cc dng hnh thi chuyn i ca chng. iu kin phng th nghim, hu ht nm pht trin trn cc mi trng tng t nh cc mi trng dng nui cy vi khun, nhng thng pH thp hn. a s nm l cc sinh vt hiu kh, nhng nm men bnh m c th pht trin trong cc giai on ngn khng c oxy. Ni chung, nm a nhit t 25 30C, mc du vy, mt s loi c kh nng gy ra cc bnh nm su, pht trin tt 37C hoc ln hn. Mt tc nhn gy bnh a nhit in hnh l loi A. fumigatus c kh nng pht trin tt nhit cao ti 50C.
Hnh 9.4. Hnh thi gi si ca Candida albicans
2.2. Phn loi nm

Da vo phng thc sinh sn bo t hu tnh ca nm hin nay ngi ta chia gii nm thnh 4 ngnh v mt nhm sau y: 2.2.1. Ngnh nm roi (Chytridiomycota) Hu ht l nm thu sinh v sng d dng, mt s c th k sinh gy cc bnh thc vt. Cc
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bo t hu tnh v c bo t v tnh u c roi v c kh nng di ng. 2.2.2. Ngnh nm tip hp (Zygomycota) L ngnh rt ph bin v thng c gi l mc bnh m. Hu ht sng hoi sinh, nhng cng c kh nhiu loi sng k sinh gy bnh ngi v thc vt. Ngoi ra trong ngnh ny c b Glomales, sng cng sinh kh ph bin vi khong 70% thc vt trn th gii. Cc bo t hu tnh l dng bo t ngh, c thnh dy, v c gi l bo t tip hp (zygospore). Cc bo t v tnh c sinh ra trong mt nang gi l nang bo t (sporangium). 2.2.3. Ngnh nm ti (Ascomycota) Ngnh ny bao gm cc loi nm si v a s nm men. Hu ht cc tc nhn gy bnh thc vt u thuc ngnh ny. Cc bo t hu tnh c sinh ra trong mt ti gi l ascus. Cc bo t v tnh c sinh ra bn ngoi (bo t trn) c gi l conidi. 2.2.4. Ngnh nm m (Basidiomycota) Cc bo t hu tnh ca ngnh ny c sinh ra bn ngoi, trn mt cu trc hnh chy c gi l m (basidium). Bo t v tnh thng khng c. 2.2.5. Nhm nm bt ton (Fungi Imperfecti hay nhm deuteromycetes) Gm cc loi nm cha bit, hoc khng c trng thi sinh sn hu tnh. Trng thi v tnh chnh l s hnh thnh conidi. Bt k mt loi nm no cha bit, hay khng c trng thi hu tnh hay qu trnh gim phn trong chu k sng ca chng, tin li, ngi ta u xp chng vo nhm ny. Hu ht cc loi ca nhm ny c mi quan h thn thuc vi ngnh nm ti v c khong 1.680 chi vi 17.000 ngn loi c m t v xp vo nhm ny. a s cc loi ca nhm ny sng trn cn, hoi sinh hoc k sinh thc vt. Mt s t k sinh gy bnh ng vt, trong c con ngi.
3. CC C CH GY BNH CA VI NM
Mc du s lng chi v loi c m t ca gii nm l rt ln (trong t in nm ca Hawksworth v cng s xut bn 1983, m t 6.000 chi vi 65.000 loi) nhng mt iu may mn l ch c khong 50 loi l c kh nng gy bnh ngi. Nm c th gy bnh thng qua 3 c ch ch yu sau y:
3.1. Gy bnh thng qua p ng min dch

Mt s nm gy ra cc p ng min dch v cc p ng ny c th dn n cc phn ng d ng (hin tng qu mn) khi xut hin cc khng nguyn nm c hiu. V d, cc loi ca chi Aspergillus sng hoi sinh rt ph bin trong t nhin l cc tc nhn gy d ng thng gp, c th gy ra cc cn co tht ph qun (asthma) v cc phn ng qu mn khc.
3.2. Gy bnh thng qua c t (mycotoxin)

Cc c t nm thng c to ra trn cc cht hu c cht khi nm ly nhim v pht trin. Mycotoxin (ch yu l exotoxin) l cc sn phm chuyn ho th cp ca nm, y l mt nhm ln cc hp cht a dng v cu trc. Nhiu cht trong s ny rt c vi ngi v ng vt khi n phi, ht phi v k c khi tip xc vi da. Chng c th gy ra hng lot cc tc ng xu i vi sc khe con ngi. Tu thuc vo lng c t v thi gian tip xc, chng c th gy ra cc bnh cp tnh hay mn tnh v c th dn n qui thai v ung th. V d in hnh nht ca c t nm l aflatoxin, tc nhn gy bnh nguy him (aflatoxicosis) ng vt ni chung v con ngi ni ring, v cc loi nm ch yu sinh c t ny thng gp trn cc c cht thc phm bo qun khng hp l, nht l cc loi ht.
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3.3. Gy bnh thng qua nhim nm k sinh (mycosis)

Nhiu loi nm c th gy ra cc bnh ngi thng qua hnh thc k sinh. S pht trin ca mt loi nm bn trn hay bn trong c th c gi l mt bnh nm (mycosis). Tu trng hp, cc bnh nm c th gy ra cc tc hi cho k ch cc mc khc nhau t v hi (lnh tnh) n cc bnh nm gy kh chu v cc bnh nng c th e do ti tnh mng ca ngi bnh.
4. BNH DO AFLATOXIN (aflatoxicosis)
4.1. nh ngha
Aflatoxicosis l bnh gy ra do con ngi v ng vt n phi thc phm b nhim aflatoxin, mt nhm cht ch yu do 2 loi nm Aspergillus flavus (hnh 9.5, hnh 9.6) v A. parasiticus sinh ra.
4.2. Phn loi v tc hi ca bnh

Tu thuc vo hm lng c t v thi gian tiu th thc phm m con ngi c th b cc bnh ng c cp tnh hay mn tnh. Bng chng v bnh do aflatoxin cp tnh ngi c thng bo nhiu vng trn th gii, ch yu l cc nc th gii th ba nh i Loan, Uganda, n , Thi Lan,... Cc hi chng c trng ca bnh l: nn ma, au bng, ph phi, co git, hn m v cht do ph no v nhim m gan, thn v tim. Cc yu t lm tng kh nng xut hin cc bnh do aflatoxin cp tnh ngi bao gm thnh phn dinh dng ca thc phm v cc iu kin mi trng thun li cho s pht trin ca nm mc trn thc phm. Ngoi ra, s qun l lng lo v yu km nh thiu h thng theo di, kim tra v x l cc loi thc phm v nng sn b nhim aflatoxin cng gp phn quan trng trong vic gy ra cc bnh do aflatoxin. Cc aflatoxin, c bit l aflatoxin B1 l cc cht gy ung th mnh trn mt s ng vt nn thu ht c s quan tm ca cc nh bnh l hc i vi cc tc ng ca chng hm lng thp trong thi gian di i vi con ngi (ng c mn tnh). Nm 1988, T chc nghin cu ung th quc t (International Agency for Research on Cancer) xp aflatoxin B1 vo danh mc cc cht gy ung th ngi. C s ca quyt nh ny da trn cc kt qu nghin cu dch t hc 2 Chu lc u, a ra c mi lin h gia ch n nhim aflatoxin v ung th t bo gan ngi. Ngoi ra, s xut hin ca cc bnh lin quan ti aflatoxin cn ph thuc vo tui, gii tnh v tnh trng dinh dng cng nh s xut hin ng thi ca cc tc nhn nguyn nhn khc nh vim gan do virus hoc cc tn thng do cc loi k sinh trng gy ra.
4.3. Phng bnh

phng nga cn bnh nguy him ny, cn bo qun tt lng thc, thc phm, khng s dng cc loi lng thc, thc phm v thc n b mc. Nu tn dng lm thc n cho ng vt nui, cn phi c kim tra x l trc khi a vo s dng. Nu cc ng vt nui n phi cc thc n nhim cc loi c t ny th ngoi cc tc hi nh gim cn, gim sn lng trng, sa,... cc sn phm chuyn ho th cp (in hnh l aflatoxin M1) ca cc c t ban u (in hnh l aflatoxin B 1) c trong sa, trng, tht vn c c tnh cao gy nguy him cho sc khe ngi tiu dng. Ngoi ra, phi c mt h thng qun l, gim st cht ch cht lng cc loi lng thc, thc phm v thc n chn nui ang lu hnh trn th trng cng nh bo qun cc kho. Cng vic ny phi c tin hnh mt cch thng xuyn da trn cc tiu chun c th v gii hn ti a cho php ca cc c t nm trong cc loi thc phm v thc n cho vt nui, k c cc loi thc n cho tm, c. C nh vy, chng ta mi c hy vng gim bt cc nguy c bnh tt do cc loi c t nm mc gy ra cho con ngi.
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Hnh 9.5. Khun lc trn mi trng Czapek ca loi A. flavus
Hnh 9.6. Cu trc sinh conidi ca loi A. flavus
5. CC BNH NM K SINH (MYCOSES)
5.1. Mt s vn quan trng ca cc bnh nm k sinh

5.1.1. Phn b Vi mt s nh cc trng hp ngoi l quan trng, a s cc loi nm lin quan ti cc bnh ngi u sng t do ngoi mi trng. Hu ht cc bnh nm mc phi u do kt qu ca tip xc tnh c nh ht phi hoc ly nhim do cy ghp m. V d, H. capsulatum tm thy trong t b ln phn di, phn chim v loi C. neoformans c mt ph bin cc chung chim b cu v cc vng t b nhim phn chim. Nhiu loi nm c tnh thch nghi theo khu vc a l ring bit, v d nh loi Coccidioides immitis tm thy cc vng kh hu sinh hc thuc Ty Nam nc M v cc vng a l tng t Trung v Nam M. Vng ny c kh hu kh hn v na kh hn, ma h nng, nc t ng bng vo ma ng v t b nhim kim. Loi C. immitis ch xut hin vng Th gii Mi (khu vc Bc, Trung v Nam M). Chnh v vy, paracoccidioidomycosis do loi
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Paracoccidioides brasiliensis b gii hn vng Nam v Trung M. Blastomyces dermatitidis, mt loi c cho l ch c pha Bc lc a Chu M, gy ra nhng dch a phng Chu Phi. Loi Sporothrix schenckii, mt tc nhn gy cc bnh nm di da phn lp c rt ph bin hoa hng, cy hong lin gai v cc thm thc vt b su mc. Qua nghin cu s phn b cho thy, loi C. albicans xut hin ming, h thng d dy rut v cc vng nim mc khc nhau nh l mt phn ca h VSV bnh thng ca c th. Loi Malassezia furfur (Pytyrosporum ovale) l nm men tm thy trn da ca ngi bnh thng, c bit vng thn, mt v da u, nhng ni giu tuyn b nhn to ngun lipid cho cc nm ny s dng. Cui cng l nhm dermatophytes gy cc bnh ringworm v athletes foot, i khi cng c tm thy trn da vng thn v u ca mt s ngi khng c cc triu chng ca bnh. Ngi ta cho rng, y ch l s xut hin nht thi ca qun th hoc mt trng thi mang ti mm bnh. 5.1.2. Xm nhim Mc min dch thin ph chng li cc loi nm gy bnh l cao tt c mi ngi. Bng chng thc t l cc trng hp nhim nm thng nh v t c gii hn. B mt da v nim mc nguyn vn l cc hng ro bo v u tin i vi s xm nhim ca nm. iu kin kh, sch, s thay th cc t bo biu m, cc acid bo v pH thp ca da c cho l cc yu t quan trng trong kh nng khng ca c th. Ngoi ra, h vi khun ca da v nim mc cng cnh tranh vi nm v ngn chn s pht trin lan rng ca nm. Tt c nhng tc ng lm thay i s cn bng ca h VSV bnh thng ca c th nh s dng corticoid, khng sinh ph rng di ngy hoc cc thay i v mt dinh dng s lm cho cc loi nm nh C. albicans pht trin mnh hn bnh thng v lm tng kh nng gy bnh. S tn thng ca cc hng ro bo v nh chn thng, ghp c quan s to iu kin cho nm xm nhp vo cc vng m lnh ca c th. Cng nh trong tt c cc trng hp ly nhim, mc nh hng i vi c th ph thuc vo c tnh ca sinh vt xm nhim, phm vi xm nhim v s ton vn trong cc c ch bo v ca c th. 5.1.3. Chn on Ly nhim nm c chn on trong cc phng th nghim bng cc tiu bn vi hc trc tip (tiu bn sinh thit hoc tiu bn t cc dch c th c nhum, soi bng knh hin vi), nui cy v chn on huyt thanh hc. Cc c im hnh thi ca VSV s tr gip cho vic nhn din tt c cc loi nm gy bnh c trong cc m ln trn mi trng nui cy. Cc yu t ny l cc k quan trng trong chn on cc trng hp nhim nm ton thn nng. Mt s cc c tnh ring bit nh cc nang bo t ca Coccidioides immitis trong m, cc t bo nm men ny chi ln in hnh ca Blastomyces dermatitidis trong cc m, cc si nm cng bo ca cc loi nm gy mucormycosis v cc t bo nm men C. neoformans c bao bc bi mt lp v (capsule) xut hin trong dch no tu (cerebrospinal fluid) v m no, gip chn on nhanh, chnh xc trong cc trng hp nhim nm su (ton thn). i vi cc tc nhn gy bnh nm c hi nh Mucor, Candida v Aspergillus, vic xt nghim vi hc cc mu bnh phm lm sng l c bit hu ch, bi ch c tiu bn vi hc nui cy c th khng tin cy, do cc sinh vt ny thng tn ti ngoi mi trng mc cao v cng c th l mt phn ca h VSV bnh thng ca c th. Cc bng chng v m bnh hc ca s ly nhim lun l yu t c gi tr nht trong chn on, mc du kt qu ca bc ny m tnh cng khng loi tr c s ly nhim nm. Cc nm gy bnh cng c th tm thy t cc m b nhim bng cch nui cy. Khi sinh vt phn lp c l tc nhn gy bnh nguyn pht nh H. capsulatum th vic chn on l chnh xc, khng th nhm ln. Vic phn lp cc sinh vt c hi nh Candida t cc khu vc b mt c th c ngha lm sng t bi kt qu ny c th l s xm nhim ca nm, nhng qu trnh nui cy cc sinh vt ny t mu mi l kt qu lun c ngha quyt nh. Nui cy cc mu bnh phm lm sng khng phi lc no cng c kt qu v s pht trin ca VSV. V d, cc trng hp nhim Aspergillus lan to cho cc kt qu dng tnh trong nui cy mu thp hn 10% cc ca xt nghim. Chnh cc nh nghin cu cng khng c cu tr li chc chn m ch gii thch rng c l ch mt phn nh ca h si c kh nng pht trin hot tnh v to ra mt chng nui cy dng tnh, hoc phi cy mt lng ln bnh phm nhim loi nm ny mi c kh nng pht trin iu kin phng th nghim. Do nm pht trin chm v thng xuyn c s chm tr ng k gia thi gian c bnh phm v kt qu nui cy dng tnh, vic da trn cc kt qu nui cy c th gy ra mt s chm tr ng k trong vic bt u liu php iu tr. Mc du c nhng hn ch nh vy nhng vic nui cy vn phi lun c
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tin hnh bi khi chng ta thc hin cng vic ny s gip cho vic chn on v iu tr tt hn. Dng khng th c hiu pht hin cc khng nguyn nm trong mt s trng hp cng rt hu ch cho chn on, c bit l cc bnh nm su. Nhiu cc test da v huyt thanh hin ang c s dng, nhng cc kt qu ca cc php th ny cng ch l bng chng c c s i vi s xm nhim. Kt lun cui cng cng phi c lm sng t di nh sng ca cc kt qu lm sng. y l mt vn kh khn trong vic lm sng t tm quan trng ca cc test da dng tnh i vi loi H. capsulatum, tc nhn gy bnh nm ton thn nguyn pht, gy ra cc dch a phng. Hu ht mi ngi ang sng cc vng ny u tip xc vi loi nm ny v to ra mt test da dng tnh, mt ch th ca tng nhy cm kiu mun. Cc test huyt thanh c nhc im ln hn trong tr gip chn on bi cc khng nguyn sn c khng c hiu. Cc test dng tnh gi c th l do s xm nhim khng c triu chng, cc trng hp ly nhim h lm sng ln trc, hoc p ng do k c c c t cc test da trc. Cc p ng m tnh gi hoc mun, c bit l bnh nhn b c ch min dch, c th cng xy ra. Gn y, cc bc pht hin cc khng nguyn nm c trong cc dch c th pht trin. Mt thnh cng nht l vic pht hin cc polysaccharid v (capsule) ho tan ca C. neoformans trong dch no tu. y l phn ng c tnh c hiu cao v nhy. Cc test tng t i vi cc loi nm khc l cha c, nhng hin ang c nghin cu trin khai a vo ng dng. 5.1.4. iu tr Mt iu cc k quan trng cn c nhn thc l khng phi tt c cc trng hp nhim nm u phi iu tr. Nh ni phn trn, con ngi c mt kh nng khng cao thin ph i vi hu ht cc tc nhn nm gy bnh. Hu ht cc trng hp nhim H. capsulatum hoc C. immitis u l cc trng hp h lm sng, t hn ch v khng pht trin thnh bnh. Mt s trng hp nhim nm Candida c p ng tt vi cc bin php tr gip nh ci thin tnh trng dinh dng ca c th ngi bnh. Cc can thip lm sng nh tim truyn, cc ng thng v vic s dng khng sinh ph rng di ngy gy nhim nm s d dng c kim sot khi cc yu t lin quan ny c pht hin v iu chnh. Hu ht cc khng sinh khng nm khi s dng ni chung u hng ti mc tiu l loi tr cc loi nm gy bnh hn l cc t bo vt ch, nhng thc t mc ch ny khng t c hiu qu nhiu. c tnh ca thuc l mt vn ln trong iu tr cc bnh nm. Ngoi ra, nhiu hp cht khng nm c hiu qu thp do nhng hn ch v tan, bn vng v s hp thu. So vi cc khng sinh khng khun th s lng khng sinh khng nm c hiu qu l rt nh, trong khi , tn s nhim nm ngy mt tng nn vic nghin cu cc tc nhn c hiu qu b sung ang c m rng. Hu ht cc khng sinh khng nm hin c c xp vo 2 loi: Tc ng ln cc mng t bo nm. Thm vo t bo v lm gin on, cn tr cc qu trnh ca t bo. Bng 9.1 di y trnh by mt s nhm v cc khng sinh khng nm ang c s dng trong iu tr.
5.2. Phn loi
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Tu theo v tr ly nhim ch yu ca nm trn c th, ngi ta chia cc bnh nm k sinh thnh cc nhm sau: 5.2.1. Cc bnh nm ngoi bin (superficial mycoses) L cc trng hp nhim nm c gii hn lp ngoi cng ca da v tc. Cc bnh ny ni chung l nh vi cc phn ng vim ti thiu hoc khng c phn ng vim, c chn on d dng v c p ng tt vi ho liu php. Nhm ny bao gm 4 bnh, hai bnh lin quan ti tc da u (black and white piedra) v 2 bnh lin quan ti vng da khng tc (tinea nigra v tinea versicolor). Nhiu ngi trong s chng ta c nm trn da v tc nhng khng c tn hiu ca bnh. i khi, cc loi nm trong h sinh vt bnh thng ca chng ta gy ra s nhim nm ngoi bin v s nhim ny khng lan su hn lp sng (stratum corneum) ca da, bnh thng nh v khng gy ra cc p ng vim v thng b b qua, khng n. Cc dng bnh ny thng thy cc vng kh hu nng, m nh nc ta. Nhng mt bnh khc l tinea versicolor (pityriasis versicolor) li thng gp cc vng n i hn l vng c kh hu nhit i. Cc bnh nm ngoi bin thng d iu tr bng cc tc nhn tiu hu keratin ti ch. Di y trnh by chi tit v mt bnh kh ph bin thuc nhm ny, l bnh lang ben (tinea versicolor). Bnh lang ben (tinea versicolor hay pityriasis versicolor) Bnh lang ben l bnh do nhim nm lp ngoi cng ca da, gy ra cc mng da mu trng hoc khng mu, c vy. Tc nhn gy bnh: Do mt loi nm men Malassezia furfur gy ra. Bnh tng i ph bin c bit l thanh, thiu nin. * Triu chng: Bnh t khi gy au hoc nga nhng c kh nng ngn cho da khi b rm nng nn to cc mng da c mu sng hn so vi vng da xung quanh. Nhng ngi c mu da en mt cch t nhin c th thy cc vt da b nhim nm sng hn. Nhng ngi c mu da trng mt cch t nhin c th quan st thy cc m da b nhim nm c mu ti hoc sng hn. Mu ca vng da b nhim nm ph thuc vo s tc ng ca nm men ly nhim i vi cc t bo sinh melanin biu b. Cc mng da ly nhim nm thng xut hin vng ngc hoc lng v c th bong vy nh. Theo thi gian, dn dn cc vt nh c th hp li thnh cc mng ln. Chn on v iu tr: Cc bc s c th chn on bnh lang ben qua v b ngoi ca vng da b nhim nm. C th s dng nh sng cc tm pht hin s ly nhim mt cch r nt. Hoc c th kim tra bng knh hin vi cc mnh biu b c co t vng da b nghi c s ly nhim nm xc nhn s chn on. Thng s dng cc loi thuc m ketoconazole bi ti ch, dung dch phun terbinafine, du gi u selenium sulfid. Du gi u ketoconazole gi trong vng 5 pht sau gi sch bng nc, dng hng ngy v lin tc trong 3 ngy. Cc thuc dng qua ng ung nh itraconazole, ketoconazole hoc fluconazole thng c p dng iu tr cc trng hp khng v lan rng. Tuy nhin, do cc thuc ny c th gy ra cc tc dng khng mong mun nn thng c xu hng thin v cc thuc s dng ti ch. Da c th khng tr li mu sc bnh thng sau khi iu tr khi bnh nhiu thng. Bnh lang ben thng hay b ti nhim. Do vy, nhiu bc s khuyn nn s dng du gi u selenium sulfid 2,5%, hoc du gi u ketoconazole hng thng phng cc trng hp ti pht. 5.2.2. Cc bnh nm da (cutaneous mycoses, dermatomycoses) Su hn mt cht trong vng biu b s bt gp cc bnh nm da nh athletes foot hay ringworm. Cc bnh ny xut hin do mt nhm nm c gi l dermatophytes gy ra. Cc bnh do nhm nm ny gy ra c th cp tnh hay mn tnh, ph thuc vo tc nhn gy bnh v tnh trng min dch ca bnh nhn. Ni chung, cc bnh ny kh iu tr hn so vi cc bnh nm ngoi bin. Cc tc nhn gy bnh l cc loi thuc 3 chi: Trychophyton, Microsporum v Epidermophyton (hnh 9.7, 9.8 v 9.9).
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Hnh 9.7. Hnh thi vi hc ca loi Microsporum nanum
Hnh 9.8. Hnh thi vi hc ca loi Trichophyton ajelloi
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Hnh 9.9. Hnh thi vi hc ca loi Epidermophyton floccosum
Cc trng hp bnh lm sng c gi l tinea v chng c m t tng ng vi v tr m c th b nhim. V d nh bnh tinea capitis lin quan ti u, tinea pedis lin quan ti chn, tinea corporis lin quan ti vng da phng thn,... Mt s loi c mt khp ni trn th gii, mt s khc b gii hn mt s khu vc a l nht nh mt s vng ca th gii. S phn b ny hin nay b o ln do nhu cu giao lu ngy mt tng ca con ngi trn ton th gii. 5.2.2.1. Phn b Nhiu loi khc nhau ca h nm da c cc sinh thi khc nhau: cc loi phn lp c thng xuyn t t c gi l geophilic (a t) mt s cc loi khc hay gp ng vt c gi l zoophilic (a ng vt); nhm th 3 l cc loi anthropophilic (a ngi), thng tm thy ngi. y l yu t rt quan trng trong vic nhn bit cc loi, xc nh ngun ly nhim c th lin quan. Cc loi nm a ngi c xu hng gy cc bnh mn tnh ngi v thng kh iu tr. Cc loi nm a ng vt v t c xu hng gy cc tn thng vim ngi nhng thng khi bnh mt cch t nhin m khng cn iu tr. Cc loi thuc nhm dermatophytes khng phi l cc thnh vin ca h VSV da bnh thng. Mc du vy chng vn thnh thong c tm thy trong cc k ngn chn ca mt s ngi. Cc loi nm ny hu nh lun gy ra cc bnh l nh mi khi chng thit lp c s ly nhim. Bnh gy ra bi cc loi nm ny gi l ringworm hoc tinea. Tn tinea bt ngun t ting Latin cp ti cc tn thng c dng hnh trn do nm gy ra da. Cc tn thng ny l c trng ca cc trng hp nhim nm dermatophytes. Tn tinea ny c dng kt hp vi tn cc phn c th b nhim nm nh tinea capitis ( u), tinea pedis (nm chn), tinea corporis (nm thn), tinea cruris (bnh nm cc np gp ca chn, tay nh bn, nch, np gp v,...). Nhng ngi khng chuyn mn thng gi tn cc bnh ny di dng cc tn khc nh athletes foot, jock itch, jungle rot,... 5.2.2.2. Xm nhim Cc nghin cu thc nghim v cc trng hp ly nhim nm dermatophytes h tr rt nhiu
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trong vic nghin cu cc biu hin lm sng ca bnh. Ngi ta cho nhng ngi tnh nguyn ngm chn vo trong nc c cha cc bo t nm gy bnh khc nhau. Kt qu h u khng b mc bnh ngoi tr trng hp chn ca h b tn thng trc khi ngm. Cc iu kin m, kn duy tr ko di cng l nhng yu t thun li cho s xut hin bnh. l cc trng hp nh i cc i giy khng l, hoc khi ph trn da bng nhng ming bng kn. Khi bnh tin trin xut hin mt phn ng vim lp biu b pha di v lp b. S bong vy thng xuyn xut hin l du hiu ca s thay biu b tng ln. Mng chn, tay, nang lng v thn tc c th b ly nhim. Cc loi nm ny c bit thch nhng m b keratin ho, chng khng xm ln su vo nhng t bo sng cng nh cc vng ca chn tc cha b keratin ho hon ton. Mc du vy, keratinase vn c tm thy mt s loi ca nhm dermatophytes. Vai tr ca n trong cc bnh do nhm nm ny gy ra vn cha c bit. 5.2.2.3. Triu chng Cc c im lm sng ca bnh ny lin quan ti s vim ca biu b, b, nang lng v tc. Yu t thc y to hin tng vim ny l phn ng min dch trung ho cc khng nguyn nm lan truyn t vng biu b b nhim nm. Qu trnh ny c th c gi l vim da do tip xc sinh hc. Tn gi kerion c dng m t dng bnh nhim nm vng da u v ru vi cc phn ng vim nng c cc mn m. M l kt qu ca cc trng hp nhim khun th cp (bi nhim vi khun). Nhim nm ton thn do dermatophytes rt him khi xy ra, v c im ny khng ph thuc vo vic vt ch b tn thng nh th no. L do chnh cho s gii hn ny ca bnh c th l s pht trin km ca nhm nm ny nhit c th ngi. Tp qun vn ho v mi trng kt hp vi cc kiu qun o, giy dp cng lm tng thm t l ca bnh nm da do dermatophytes. Cc nghin cu c tin hnh trn cc qun th dn c v cc gia nh c tp tc ring cho thy, cc iu kin sng vi mt dn c ng c, gn nhau l cc yu t quan trng trong s lan truyn ca bnh. Cc yu t min dch cng gy nh hng ti t l ca bnh v cc kt qu nghin cu cho thy min dch qua trung gian t bo c vai tr quan trng i vi s ly lan ca bnh. Cc iu tra khng y t cc nghin cu dch t hc v cc bo co ca cc ca bnh cho thy nhm nm ny l mt trong s cc loi nm ph bin nht gy ra cc bnh ngi. Cc bnh nm da do nhm ny gy ra l trong s cc bnh da ph bin nht tr em di 12 tui v l bnh da ph bin th 2 tui ln hn. Cc nhm tui khc nhau b nhim cc bnh nm ny cc vng khc nhau ca c th. Cc i tng thuc khoa nhi, bnh nm vng da u l ph bin nht v bnh chim t l cao nht tui t 5 10. Sau tui dy th, cc bnh ny t ph bin. L do ca s thay i ny vn cha c bit chnh xc, nhng c th do s thay i trong thnh phn ca b nhn, s thay i bt u xut hin tui dy th, c bit tng s cc acid bo bo ho. Cc acid ny c hot tnh khng nm t nhin. Mt khc bnh athletes foot him khi xut hin tui th u, nhng dn dn bnh cng ly nhim ph bin v cn li sut c cuc i. iu ny c th l do ngi ln thng mang giy nn c mt t l cao ca bnh nm k chn (athletes foot) do nhm nm dermatophytes gy ra. M ngi ta thy c mt t l thiu cn xng ca bnh nm da u (ringworm of the scalp) tr em da en nhng cc nh nghin cu vn cha tm c nguyn nhn. Nhng ngi bn x n c t l mc cc bnh nm ny cao hn so vi ngi Chu u n nh c. Cc nghin cu t cuc chin tranh Vit Nam cho thy rng, bnh athletes foot cc qun nhn M ch yu l do loi Trichophyton mentagrophytes gy ra. Tri li, ngi Vit Nam chng ta li nhy cm hn vi loi Trichophyton rubrum. T l bnh nm do dermatophytes gy ra nam cao hn n: cc bnh nm vng da u do nhm nm ny gy ra c t l gia nam v n l 3 : 1, ca bnh nm bn chn, k chn l 6 : 1. Cc bnh vng bn v sinh dc cng ph bin hn nam gii v him gp n. Nhim nm mng tay ph bin hn n gii, nhng nhim nm mng chn li thng xuyn lin quan ti nam gii. 5.2.2.4. Chn on
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Kim tra vi sinh hc cc mnh vn c co ra t b mt da cc vng b nhim nm tm nm. Cc khun lc ca dermatophyte c cc c tnh hnh thi phc tp. Cc c tnh ny gip phn bit cc chi v loi. Di knh hin vi chng khc nhau c im hnh thi ca cc i bo t a bo c trng (macroconidi). 5.2.2.5. iu tr Cc bnh nm do dermatophytes c iu tr ti ch v ton thn. iu tr ton thn l cn thit khi tc v mng b nhim nm bi cc thuc c iu tr ti ch khng xm nhp c vo cc m ni nm tn ti v pht trin. Cc dng thuc iu tr ti ch khc nhau nh kem, thuc m, thuc ra v phn nc (l ch phm lng dng p trn da hay nim mc, thng cha cc cht lm se, n mn v gim au) c iu tr thng xuyn trong vng 3 tun hoc hn s lm mt ht cc vng da nhim bnh ringworm ti ch. Cc cuc th nghim c gim st khng chn c mt cht tt nht trong s cc thuc chng nm c s dng. Hai tc nhn chng li bnh ringworm c dng iu tr ton thn bng ng ung, l griseofulvin v ketoconazole. 5.2.3. Cc bnh nm di da (subcutaneous mycoses) Cc bnh nm di da l mt nhm bnh ring bit lin quan ti b (dermis) v cc m di da, hoc cc cu trc m k tip. Tc nhn gy bnh l cc loi nm phn lp c ph bin ngoi mi trng v ch gy ra bnh khi c iu kin thun li nh: b cn trng t, try xc da do cc nguyn nhn khc nhau nh gai hoc cc mnh v,... Nhm ngi c nguy c cao i vi cc bnh ny l ngi bun bn hoa, nng dn, ngi hay tip xc vi c,... Nhiu trng hp b bi nhim nm khi mc cc bnh do vi khun. Cc bnh ny t gp v ch gii hn cc vng nhit i. Bnh pht trin chm v thng c xu hng ko di. Mt v d in hnh ca nhm bnh ny l bnh do nm Sporothrix schenckii gy ra. y l mt loi nm lng hnh c th chuyn thnh dng nm men khi nui cy 37OC trn cc mi trng nui cy giu dinh dng phng th nghim hoc khi k sinh ngi. Bnh ny trc y ph bin Chu u nhng hin nay t gp. Hin nay bnh xut hin ch yu Chu M, Nam Phi v c. Triu chng bnh thng gp giai on u l mt tn thng lot v c th pht trin dn n vim cc mch bch huyt. Cc bnh nm di da ni chung khng p ng tt vi ho liu php. Trong nhiu trng hp thng phi ct b tn thng hoc phu thut kim sot s ly lan. Ring bnh do Sporothrix schenckii (sporotrichosis) l mt ngoi l, bnh p ng tt vi KI hoc ketoconazole qua ng ung. 5.2.4. Cc bnh nm ton thn Cc bnh nm ton thn (systemic mycoses) l cc ly nhim nm su vo cc c quan bn trong c th. Cc bnh nm ny c th do cc tc nhn gy bnh nguyn pht nh H. capsulatum hoc Coccidioides immitis, chng tn cng nhng ngi khe mnh. Hoc do cc loi nm c hi nh Candida albicans, chng ch gy bnh khi c cc iu kin thun li v ni chung khi vt ch b suy nhc, suy gim h thng min dch, cc loi nm ny s tng cng xm nhim v gy bnh. Trong cc bnh nm ton thn, nguyn pht, hu ht cc i tng kho mnh, c cc du hiu bnh nh hoc cc biu hin h lm sng. Ngc li, cc trng hp nhim nm c hi cc bnh nhn b suy nhc th hu nh u b mc cc bnh nng. 5.2.4.1. Cc bnh nm ton thn nguyn pht Mt dng ca cc bnh nm ton thn do cc tc nhn gy bnh ban u gy ra Histoplasma capsulatum: + L mt sinh vt c trong t, s pht trin ca n tng ln khi c cc loi phn di, phn chim. + Gy histoplasmosis khu vc trung tm min Ty nc M, c gi l histobelt. + Chuyn thnh dng nm men khi tn ti nhit trong c th. + Xm nhp i thc bo v khng li s thc bo nu phm vi xm nhim rng hoc cc i thc bo khng hot ng. + Bnh do Histoplasma tng t bnh lao (tuberculosis). Cc sinh vt c th duy tr s sng c lp trong phm vi cc tn thng b vi ho, v v th c kh nng ti nhim. + Histoplasmosis tin trin cc bnh nhn b suy gim min dch.
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Cc bnh nm ton thn gy bi cc tc nhn ban u khc: + Blastomycosis ng h hp do Blastomyces dermatitidis. + Coccidioidomycosis do Coccidioides immitis. + Paracoccidioidomycosis do Paracoccidioides brasiliensis. 5.2.4.2. Cc bnh nm ton thn c hi a) Mt s c im ca cc loi nm gy bnh c hi Bnh thng cc loi nm c hi khng gy bnh, ch ly nhim v gy bnh ngi c h thng bo v b tn thng, c bit l cc i tng b gim s lng cc bch cu a nhn trung tnh, lm tng mn cm vi cc bnh nhim trng. b) Cc loi nm gy bnh c hi ch yu Candida albicans l mt loi c trong h VSV bnh thng ca c th. Aspergillus fumigatus v cc loi ln cn: Cc tc nhn gy bnh c ph bin ngoi mi trng, gy bnh lan to cc bnh nhn b suy gim min dch nng. Cryptococcus neoformans, mt tc nhn gy bnh vim mng no ph bin bnh nhn AIDS. Mt s loi khc thuc chi Penicillium (Penicillium marneffei) v nhm Zygomycetes nh Mucor, Absidia v Rhizopus. c) Kh nng gy bnh Cc bnh nhn c nguy c cao nhim nm c hi bao gm cc i tng c cc khi u c tnh, cy ghp c quan hoc tu xng, bng, tn thng v b cc bnh cn s dng cc ng thng tnh v ng mch di ngy. Bnh nhn tri qua cc ca phu thut, s dng liu php khng sinh ph rng v h VSV bnh thng ca h c du hiu suy gim. Trong cc trng hp nh vy, cc loi nm nh Candida albicans s pht trin khng kim sot c v c th thay th v tr ca cc vi khun khc trong h VSV bnh thng v chuyn thnh dng gy bnh. Qua kinh nghim lm sng, s gim s lng bch cu chc nng a nhn trung tnh l yu t quan trng nht nh hng ti p ng ly nhim nm c hi. Nhng trng hp ny l kh iu tr ngi bnh thng, hu nh khng c kh nng cu cha ngi c t bch cu chc nng. Tr em m yu, non v ngi gi cng rt kh cu cha khi mc cc bnh nm ny. Cc loi nm gy ra cc bnh c hi hu nh c mt khp ni. Chng c th l mt phn nm trong h VSV bnh thng ca chng ta, hoc b c th chng ta ht phi hay n vo t ngoi mi trng. Bng 9.2 di y tm tt c im ca cc bnh nm c hi thng gp.
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Mt s bnh nm c hi thng gp:
5.3. Cc bnh nm ton thn c hi thng gp

5.3.1. Bnh do Cryptococcus 5.3.1.1. Mt s nt v chi Cryptococcus C khong 37 loi trong loi C. neoformans gy bnh ngi, mc v pht trin 37OC. Cc loi khc: C. anbidus, C. laurentii, C. terreus, C. uniguttulatus phn lp c mi trng ngoi (khng kh, nc v t), sng hoi sinh ngi (da, ph qun), pht trin chm 37OC. 5.3.1.2. Loi C. neoformans 1894, Sanfelice phn lp ln u tin t nc o vi tn gi Saccharomyces neoformans. 1894, Phn lp ln u tin ngi t mt vt lot chn vi tn cng thuc chi Saccharomyces. + 1901, Wuillemain xp loi ny vo chi Cryptococcus. + 1914, xut hin ca u tin b bnh vim mng no do Cryptococcus. cp ti 2 th: C. neoformans var. neoformans v C. neoformans var. gattii. 5.3.1.3. Dch t hc Th C. neoformans var. neoformans c mt khp ni, phn b rng ri ngoi mi trng. Ngun bnh chnh l phn chim, trong c bit l chim b cu. Nm c th sng c 2 nm trong t. Th C. neoformans var. gattii: 1990 phn lp ln u tin c t cy bch n. Phn b nhng vng nhit i v cn nhit i (Chu c, Chu v California M). Him thy trng hp bnh gy ra do th ny nhng ngi lm vic trong rng bch n. 5.3.1.4. Phng thc ly nhim Do ht phi t bo nm l ch yu, bnh cng c th xy ra do chn thng. 5.3.1.5. Kh nng gy bnh
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L bnh nm c hi, thng gp nhng ngi b suy gim min dch, Bnh nhn b HIV l ch yu (khi t bo CD4 < 200/mm3, l nhng t bo tr gip T, chng l mc tiu i vi s ly nhim HIV). Ngi s dng liu php corticoid, bnh t min. Cy ghp c quan (c bit l thn thng d mc), b u lympho, Hodgkin, u tu. Nm ny thng gy bnh phi, no, da, xng, mt, tuyn tin lit. Bnh do C. neoformans phi: Ln u nhim thng khng c triu chng hoc st nh, ho, kh th, au hng. Nu suy gim min dch bnh s pht tn rt nhanh. Bnh do Cryptococcus no v mng no: Bnh pht trin t t, xut hin hi chng mng no; du hiu in hnh l au u, st, bun nn, nn, cng gy v s nh sng. Ri lon tnh cch, tr nh, bun ng, hn m v co git. U do nm (t gp): khi cng, c nh. Bnh do Cryptococcus da: Xut hin hch nh, ban , cng lm, apxe, dng khng in hnh ging mt molluscum hoc herpes, bnh ch yu mt v chn. * Bnh nhng v tr khc: + Xng: Gy apxe lnh (au). + mt: Vim li vng mc, vim mng sng. + gan: Gy vim gan. + Mt s v tr khc nh tim, thn, tuyn tin lit, xoang, thc qun,... 5.3.1.6. Tin lng Ph thuc vo chn on sm hay mun, nu chn on sm, iu tr kp thi, bnh s khi. 5.3.1.7. Chn on + Bnh phm: Dch no tu, nc tiu, sinh thit, cy mu. + Soi trc tip: Dng t bo nm men hnh trn, ng knh 3 12m, ny chi nhiu pha. Khi nhum vi mc tu pha long, carmin, nhum bng Giemsa nhn thy r nang (capsule). + Cy nm: 37OC nm mc sau 2 5 ngy (i khi lu hn) nhy cm vi actidione. Xc nh bng mt thng: Khun lc c mu kem, sng, nhy v trn. Lc u c mu trng sau chuyn sang mu t. Xc nh VSH: Dng nm men hnh cu, c nang (capsule) cy trn mi trng Plate Count Broth khng c si. Nhy cm vi actidione. Phn ng ure dng tnh 37OC. Khng ln men ng, kh yu cc mui tetrazolium, ng ho inositol. Chn on theo loi: Phn ng ure dng tnh (trc 4h) phn ng phenol oxydase. 5.3.1.8. M hc V capsule (bn cht polysaccharid) ch c nhum mu bng muxicarmin, nu khng nhum carmin th ch xut hin cc vng sng. 5.3.1.9. iu tr Trng hp nng: Amphotericin B: Tim tnh mch vi liu 0,7 1mg/kg/24h, kt hp vi 5fluorocytosine bng ng ung hoc tim tnh mch liu 150mg/kg/24h (nu bnh nhn dng tnh vi HIV, dng liu 100mg/kg/24h v iu tr t 15 ngy cho n 6 tun). Sau , tip tc iu tr bng fluconazole bng ng ung hoc tim tnh mch vi liu 400mg/24h (c th ti 800mg/24h) trong thi gian 8 tun. Trng hp nh: Fluorocytosine 400mg/24h kt hp vi 5fluorocytosine 150mg/kg/24h hoc itraconazole 600mg/24h trong thi gian 7 ngy, sau dng 400mg/24h. Thng kt hp vi 5fluorocytosine. Dung np v c tnh ca thuc:
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+ Amphotericin B: Dung np km, nhc u, rt run, vim tnh mch huyt khi. + Fluconazole: Dung np tt, c tnh vi gan v h tiu ho. Cc trng hp phi hp thuc khng nn s dng: + Amphotericin B: * Thuc c vi thn: nhm aminozide, cyclosporine. * Thuc tc ng ln kali huyt: corticoid, digitan, diure, cc thuc chng hen, thuc lm tng ti hp thu. + Fluorocytosine: Trnh phi hp vi thuc chng gim bch cu. + Fluconazole: Sulfamid, thuc chng ng ng ung, cyclosporine. iu tr d phng: Tip tc iu tr trong tnh trng suy gim min dch ko di: fluconazole 200mg/24h. 5.3.2. Bnh do Aspergillus (aspergilloses) 5.3.2.1. Cn nguyn Bnh do cc loi thuc chi Aspergillus gy ra. c bit l cc loi Aspergillus fumigatus, A. flavus, A. niger, A. nidulans. Ch yu l do ht phi bo t ca cc loi nm ny. Ngoi ra, c th nhim bnh do dng c phu thut, do tip xc trc tip vi cc loi nm ny. Chi Aspergillus c khong trn 200 loi, c mt khp ni, sng hoi sinh trn cc c cht cht trong t nhin, trong t. S pht trin hoi sinh ca chng to ra hng triu bo t pht tn trong khng kh, trong bi nh, my iu ho nhit , my ht m, cc loi giy, ba carton,... Ngoi vic gy bnh bng 2 c ch: gy d ng v gy ng c bng c t, cc loi ca nhm ny cn gy cc bnh nguy him bng kh nng xm nhim cc c quan ca c th. 5.3.2.2. Cc yu t thun li cho s xm nhim ca nm y l cc loi nm c hi, chng ch pht trin trn ngi khi gp cc iu kin thun li sau: Cc yu t chung: + Gim bch cu ht. + Suy gim min dch. + S dng liu php corticoid. Yu t ti ch: Tn thng ph qun. Yu t mi trng: Tn ti mt bo t trong khng kh v con ngi ht phi vo phi. 5.3.2.3. Cc bnh do Aspergillus gy ra Cc bnh ng h hp: Ch yu do ht phi bo t. U nm do Aspergillus xut pht t cc hang lao, apxe, dn n phi, hoi t, ung th. Dnh mng phi do nm xm nhim (ch yu bnh nhn suy gim min dch trm trng) m biu hin r nht l gim bch cu a nhn trung tnh. Bnh nhn st, kh th, ho, au hng v bnh pht tn nhanh ln no, tim, da,... Vim xoang do Aspergillus: * Cc bnh cc b phn su trong c th: Thng do phu thut nh vim mng bng, vim xng, tu. Nhim nm vo no thng do phu thut thn kinh, vin mng trong tim do phu thut tim,... * Cc bnh cc b phn bn ngoi c th: Bnh nm tai: Bi nhim vim tai ngoi do dng khng sinh hoc corticoid. Vim mng: Lan su di mng. Nhim nm da: nhng ngi b bng. Nhim nm mt: Vim mng sng, gic mc do tn thng mt. 5.3.2.4. Tin lng Din bin xu trong cc trng hp suy gim min dch trm trng, t l t vong trong cc dng bnh: 90% trong ghp xng; 60 80% ghp c quan khc.
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5.3.2.5. Chn on C th bng cc phng php: Xc nh nm, Xquang v huyt thanh hc. Ly bnh phm: phi: ly m, dch ph qun cc v tr khc nh dch xoang, dch ng tai ngoi, sinh thit. Soi trc tip: Si nm c kch thc bnh thng sau 7 ngy, ng knh si 2 3m, phn nhnh to gc nhn so vi si ban u, cc khi conidi hnh cu hoc to trn. Thng trong soi trc tip bnh phm phi va c bo t, va c si nm mi c gi tr trong chn on. Nui cy: Mi trng Sabouraud (SDA) 37OC v 27OC. A. fumigatus c th pht trin tt nhit cao ti 57OC. A. flavus c th pht trin tt nhit cao ti 42OC. Nm mc sau 1 n 3 ngy, nhy cm vi actidione. + Xc nh i th: Khun lc mi mc c mu trng, sau chuyn mu do s xut hin ca bo t, c th c mu nu, en, xanh l cy hay mu vng. Mi trng chun dng nui cy xc nh l Czapek. + Xc nh vi th: Cc khi conidi hnh ct hoc hnh tia. * Conidiophore: Trn hoc x x (c gai), c mu. * Bng (vesicle): Hnh cu hoc gn cu. * Th bnh (phialid): Hnh chai, 1 lp hoc 2 lp (khng c hoc c cung th bnh). * Conidi hnh cu, trn hoc sn si. 5.3.2.6. iu tr Amphotericin B: Tim tnh mch 1mg/kg th trng/24h, l thuc c ph hot tnh rng, dung np tc thi, gy cm gic n lnh, kh chu, st, vim tnh mch huyt khi, au tnh mch, c vi mu v thn. Cn theo di cng thc mu v chc nng gan, thn. Itraconazole: Ung: 200 400mg/24h. Ph hot tnh rng vi c nm men, nm si v nm lng hnh. Dung np tt, gy kh chu ng tiu ho (1 4%), ni ban, nga. Voriconazole: Ph hot tnh rng vi c nm men, nm si v nm lng hnh. Dung np tt, cc trng hp ri lon r rt chim 30%. Gy ri lon cc test v gan (10%), ban m ay (5%). + ng dng: Ung hoc tim tnh mch. Ung 400mg/24h chia 2 ln. Tim tnh mch 6mg/kg th trng chia 2 ln/24h vi ngy u, sau 4mg/kg th trng chia 2 ln/24h. Caspofungine: Tim tnh mch 70mg/24h vi ngy u, sau 50mg cho 24h. L thuc khng nm ph rng vi c nm men, nm si v nm lng hnh. + D phng: Trnh ht phi bo t, c bit vi ngi c hi chng AIDS, n ung sch, sch, kim sot nm tt trong mi trng khng kh v thc phm, khng trng cc cy hoa trong phng. 5.3.3. Bnh do histoplasma (histoplasmosis) 5.3.3.1. Cn nguyn Do cc loi nm thuc chi Histoplasma gy ra. Trong , ng ch l loi H. capsulatum vi hai th quan trng l H. capsulatum var. capsulatum v H. capsulatum var. duboisii. 5.3.3.2. Dch t hc Th H. capsulatum var. capsulatum phn b khp ni trn th gii, lc xut hin nc ny lc xut hin nc khc. Cc vng dch ch yu l: Chu M c bit l Hoa K thung lng 2 con sng Ohio v Mississipi; Mehico; Trung M; vng Caribe; Brazin; Achentina,... Cc vng rng su thuc Chu : n , Sri Lanka, Philippine, Nht Bn, Thi Lan, Vit Nam,... Chu Phi: Nam Phi. Chu i Dng: Newzealand. Chu u.
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5.3.3.3. Phng thc ly nhim Ch yu l do ht phi bo t. + Nm thng xut hin trong t c nhiu nit. + Trong phn ng vt nh phn chim, phn di, him khi ly qua ng tiu ho hoc qua da try xc, khng ly t ngi sang ngi. 5.3.3.4. Kh nng gy bnh * Vi ngi c h thng min dch tt: Nu b ly nhim t s khng c triu chng bnh (90%). Nu nhim nhiu nh dng phi thng lnh tnh. Thi gian bnh 5 20 ngy. Vi cc hi chng gi cm, st, ho khan, i khi kh th, ho ra mu, au hng, au c, tip theo l s vi ho phi. * Vi ngi b suy gim min dch: Rt d b ly lan ton thn, i khi tn ti lu sau khi nhim vi cc triu chng st, suy nhc nghim trng, gy i, gan to, lch to, xut hin hch, lot ming, hu. Da b thng tn, xut hin cc nt sn, ni hch, pht ban. Cc bnh ni tng thng ng tiu ho gan, thn, tim,... C a nhng ngi d b mc bnh: + ngi c HIV dng tnh: Thng b mc khi s lng t bo CD4 khong 200/mm3. + nhng ngi khng b HIV thng mc khi b: Ung th, tiu ng, nghin ru, iu tr corticoid di ngy. * V cc yu t sinh hc: Thiu mu, gim tiu cu, tng men gan. 5.3.3.5. Chn on H. capsulatum var. capsulatum l loi nm lng hnh. Khi cy 25OC c dng si (trng hp hoi sinh), dng nm men khi cy 37OC (trng hp k sinh). Ly mu: phi ly m. Ly mu, tu xng, sinh thit (da, hch, gan). Soi trc tip: C dng nm men: hnh cu, hnh trng kch thc 13 2,5 m, thng ny chi mt pha. Khi nhum bng giemsa (MGG) hoc xanh metylen xut hin vng sng xung quanh t bo (capsule). Nui cy xc nh: Mu ly phi tin hnh th ngay khng ly nhim vi khun, vi nm. Xc nh i th (bng mt thng): * Trn mi trng Sabouraud (SDA) 25 27OC: khun lc lc u hon ton c dng si, sau chuyn sang dng bt (do xut hin nhiu bo t). T mu trng chuyn sang mu be ri nu nht. Mt tri khun lc khng mu (i khi c mu vng cam). Lu : Phn bit vi nhng khun lc mc do nhim t khng kh. * Trn mi trng thch tim no thm 5% mu 37OC: khun lc dng nm men, c mng v cc np gp x x. T mu trng chuyn sang mu be ri nu nht. + Xc nh vi th: * Trn mi trng Sabouraud 25 27OC: Si nm sau 7 ngy mnh, trong sut, c ng knh 2 3m. Microconidi trn hoc hi sn si, hnh trn hoc qu l, ng knh 2 3m thng c mang bi mt cung. Macroconidi: trn, thnh dy, c nm, kch thc 8 16m, c mang bi mt cung nh bo t di (cy di ngy). * Trn mi trng thch tim no c 5% mu 37OC: cc bo t ny chi nh, ng knh 2 5m, hnh trn hoc hnh trng ny chi gc hp. 5.3.3.6. iu tr Vi cc dng vim phi cp v dng pht tn nhng ngi suy gim min dch, vic iu tr gm hai giai on: iu tr tn cng v iu tr duy tr. + Amphotericine B (tim tnh mch): 1mg/kg th trng/24h.
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+ Itraconazole (ung): 400mg/24h (vi ngi b suy gim min dch). 200mg/24h (vi ngi c h min dch bnh thng). + Ketoconazole (ung) 400 600mg/24h (c th tng ti 800mg/24h) trong 3 thng. Sau dng liu 400mg/24h trong sut 6 12 thng. Kt hp vi fluconazole (ung hoc tim tnh mch) 400mg/24h. Lu : Thuc c vi mu, gan v thn, cn kim tra cng thc mu, chc nng gan, thn. 5.3.3.7. D phng + Phng trnh ht phi bo t (c bit cc a im nh hang, ng, chung tri, g, vt,...) + Dng: Itraconazole 200mg/24h. i vi th Histoplasma capsulatum var. duboisii ch yu xut hin Chu Phi, Vit Nam cha bt gp trng hp no. 5.3.4. Penicilliosis v loi Penicillium marneffei 5.3.4.1. Tc nhn gy bnh Bnh do cc loi thuc chi Penicillium (hnh 9.10) gy ra. Chi ny c khong 200 loi, trong , ch c loi P. marneffei l gy bnh trn ngi. Ch yu l cc i tng b suy gim min dch c ngun gc ng Nam hoc tng sng v hot ng ng Nam . Loi ny mc tt 37OC, cc loi khc sng hoi sinh trn cc c cht cht, mc km hoc khng mc 37OC v rt ph bin ngoi cnh.
Hnh 9.10. Hnh thi vi sinh hc chi Penicillium
5.3.4.2. Dch t hc Vt ch thng gp l chut, thng phn lp c t cc mu t cc hang ca cc loi gm
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nhm ng Nam nn cc vng dch a phng ch yu ng Nam , ni c nhiu ngi mc hi chng AIDS nh Bc Thi Lan, Nam Trung Quc, Vit Nam, Campuchia, Malaixia, Hng Kng, i Loan,... 5.3.4.3. Phng thc ly nhim Qua ng hng khng, du lch,... Qua da do chn thng, tai nn phng th nghim,... 5.3.4.4. Kh nng gy bnh Nm pht tn trong ton b h li ni m vi triu chng lm sng a dng, mc ly lan nhanh v mc nguy kch ph thuc vo hin trng min dch ca c th ngi bnh. Vi ngi nhim HIV b nhim P. marneffei s xut hin bnh khi t bo D4<50/ml mu. Vi ngi c HIV m tnh: Do s dng liu php corticoid di ngy, b bnh t min. Thi k bnh: C th ti 11 nm. 5.3.4.5. Triu chng Giai on u st, ho dai dng, st cn, xut hin hch, ph i lch. Giai on k tip l tn thng da, di da, cc nt nh hoc cc mn to, cc hch dng trng c mt, ton thn, cc u ca ngn tay v chn. Gy lot h tiu ho ni tng gy vim mng ngoi tim, vim mng bng, vim mng phi. 5.3.4.6. Tin lng Nu khng iu tr s t vong; iu tr sm 80% trng hp c cu sng. Bnh pht trin nhanh v trm trng hn nhng bnh nhn b SIDA. 5.3.4.7. Chn on Chn on phn bit: Lu trnh nhm ln vi cc bnh nm ni tng do Histoplasma, bnh nm da do Cryptococcus gy ra. Chn on tm nm: + Ly mu bnh phm: Da, tim, dch no tu, tu xng, sinh thit t chc, cy mu. + Soi trc tip: Trc ht lu P. marneffei l loi nm lng hnh: c dng si khi cy 25OC (dng hoi sinh), dng nm men khi cy 37OC (dng k sinh gy bnh). Nhum mu bng MGG (MayGrunwaldGiemsa), t bo dng nm men c dng hnh trn hoc hnh tr, kch thc 3 2 5m, c vch ngn gia v khng c chi. + Nui cy nm: Nui cy 25OC v 37OC. Nm mc sau 2 5 ngy trn tt c cc mi trng thng dng, nhy cm vi actidione (cycloheximide, mt loi thuc khng nm do mt loi x khun l Streptomyces griseus sinh ra). Xc nh i th: Mi trng Sabouraud hoc SDA 25 27OC: Khun lc nm c dng si, lc u c mu vng, sau l nu, xanh chuyn sang nht v cui cng chuyn mu ti. Mu lan to khp mi trng nui cy. + Xc nh vi th: Mi trng Sabouraud hoc SDA 25 27OC, conidiophore trn nhn v phn nhnh, cung th bnh (metula) c t 3 5 v chiu di khng u nhau. Th bnh ngn, rng, thng ng, s lng t 4 6. Conidi trn, hnh van v to thnh chui ngn. Mi trng Sabouraud hoc SDA 37OC, chuyn t dng nm si sang dng nm men trong 2 tun. Khun lc chuyn sang mu trng c, trn bng. T bo hnh cu hoc van, ng knh 3 7m (nhn ln bng cch phn chia). 5.3.4.8. iu tr iu tr tn cng trong 2 4 tun: + Cch 1: Tim tnh mch amphotericin B (0,6mg/kg th trng/24h) kt hp vi 5 fluorocytosine 100mg/kg th trng/24h, c th s dng voriconazole. + Cch 2: iu tr duy tr 2 6 thng:
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* Itraconazole 400mg/24h. * Ketoconazole 400mg/24h. * Fluconazole 400mg/24h. iu tr d phng th cp: Do bnh thng ti pht nhng ngi b AIDS (25%) nn sau t iu tr tn cng, thng phi dng liu d phng itraconazole 200mg/24h sut i.
CU HI LNG GI
Hin tng lng hnh gp cc loi nm no sau y? A. Sporothrix schenckii. B. Aspergillus parasiticus. C. Histoplasma capsulatum. D. Penicillium marneffei 2. Aflatoxin c sinh ra bi A. Penicillium notatum. B. Crypyococcus neoformans. C. Aspergillus flavus. D. Fusarium moniliforme. 3. Nhm nm bt ton bao gm nhng loi nm cha tm thy trng thi sinh sn hu tnh. A. ng. B. Sai. 4. c im no sau y khng ng trong bnh lang ben? A. Vi nm gy bnh a cht bo. B. Vi nm gy bnh a keratin. C. Hay m hi l yu t thun li cho bnh. D. Cc mng da bnh thng sng ty ln. E. n Wood l phng tin chn on v theo di bnh tt nht. 5. V i tnh vi k ch, ngi ta chia vi nm gy bnh da ra 3 nhm A......................... B. ........................... C. ............................. 6. U nm do Aspergillus hay gp ngi c hang lao hay dn ph qun. A. ng. B. Sai. 7. Khi mun tm Histoplasma capsulatum, ngi ta phi x l bnh phm trong vng........................, l do l......................................................... 8. Ngi ta c th gp bnh do vi nm Aspergillus di dng A. bnh d ng. B. bnh lan to. C. vim ng tai ngoi. D. bnh phi. E. tt c cc th trn. 9. Vi nm Cryptpcoccus neoformans c rt t trong rut chim b cu, ch thc s pht trin mnh khi theo phn ra ngoi cnh. A. ng. B. Sai. 10. Dng nm men ca Penicillium marneffei sinh sn ch yu bng cch 1.
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A. ny 1 chi. B. ny 2 chi. C. ny nhiu chi. D. phn i bng vch ngn. 11. ng vt mang nm Penicillium marneffei l A. Ch. C. G. E. Chut ti. B. Mo. D. Chut tre.
PHN BN
Chng 10
I CNG V VIRUS
1. I CNG
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Trc Cng nguyn, du hiu v bnh bi lit, bnh di, bnh u ma c pht hin. Tuy nhin thi nhn loi cn cha bit c cc nguyn nhn gy ra nhng cn bnh nguy him y. Sau thi gian rt di, mi n nm 1884, Louis Pasteur chng minh mm gy bnh di c th ly truyn v c th nui cy bng cch tim truyn qua ng vt thc nghim, v nm 1885, ng iu ch c vaccin phng di. Nm 1898, M.W. Beijerinck gi mm bnh khm thuc l l cht dch c hot tnh truyn nhim, v gi bng ting Latinh l Virus (mm c). Thut ng virus c s dng t y. Nm 1935, W.M. Stanley ln u tin phn tch v kt tinh c virus khm thuc l (TMV = Tobaco mosaic virus). Nm 1970, D. Baltimore v H.M. Temin (chung gii Nobel, 1975) pht hin ra enzym phin m ngc (Reverse Transcriptase) trong cc virus ch c mt si n ARN, m ra tng lai mi m cho k thut di truyn trong sinh hc phn t. Do s pht trin trong nghin cu v virus m t trc n nay c nhiu nh ngha khc nhau v virus, di y l khi nim tng i r v virus: Khi nim: Virus l nhm VSV cha c cu trc t bo, v cng b nh, khng quan st c bng knh hin vi thng, ch cha mt loi acid nucleic, k sinh bt buc trong cc t bo sng iu khin h thng trao i cht ca t bo ch m sao chp acid nucleic, protein,... ri tin hnh lp ghp trng thnh, sinh sn. Trong iu kin ngoi c th, virus c th tn ti lu di di dng i phn t ho hc khng sng v c th truyn nhim. Nh vy virus c 3 c im sau: Virus c kt cu i phn t sinh hc cha c cu to t bo, khng c hin tng sinh trng c th. Tn ti s chuyn bin tng h gia dng VSV k sinh chuyn bit trong t bo sng v dng phi sinh vt bn ngoi t bo. Mi loi virus ch cha mt loi acid nucleic, hoc ch ADN hoc ch ARN.
2. HNH THI V CU TRC CA VIRUS
2.1. Kch thc v hnh thi ca virus
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Kch thc ca virus rt nh b, c th lt qua cc nn lc vi khun, chnh v th m khng th quan st c qua knh hin vi quang hc. Kch thc virus thng c o bng nanmet (1nm = 109m). S dng knh hin vi in t v cc k thut ph tr c th quan st, o c, nghin cu t m hnh thi ca tng loi virus (hnh 10.1). V hnh thi hc virus c hnh dng kh phong ph: dng hnh que, hnh khi a din v t hp phc hp ca cc dng (hnh 10.2).
2.2. Cu trc ca virus

Nh nu, virus cha c cu to t bo, mi virus khng th c gi l mt t bo m ch c gi l ht virus hay virion (virus particle). Virion l mt virus trng thnh, c kt cu hon chnh vi thnh phn ch yu l acid nucleic (ADN hoc ARN) c bao quanh bi lp v protein. Acid nucleic ht nhn nm gia ht virus, l h gen ca virus, cn protein bc bn ngoi to thnh v gi l capsid. Capsid c to thnh bi cc tiu phn capsid cn c gi l capsome, mang cc thnh phn khng nguyn (KN) c th v c tc dng bo v li acid nucleic. Li v v hp li to thnh nucleocapsid, l cu trc c bn ca virus.
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Mt s virus c cu trc phc tp hn: Bn ngoi capsid cn c mng bao lipid hay lipoprotein, c th cn c cc mu gai (spikes) bm xung quanh. Mng bao thc cht l mng t bo cht ca t bo ch, nhng c virus cu to li v mang tnh KN c trng ca virus. Mng bao c th b cc dung mi ho tan lipid ph hu (ete, aceton,...) (hnh 10.3).
Cc kt qu nghin cu s dng k thut nhiu x rnghen, knh hin vi in t cho thy cc virion c 3 dng cu trc chnh l cu trc xon, cu trc i xng khi, v cu trc hn hp vi cc c trng rt c th. 2.2.1. Virus cu trc xon Cc virus khm thuc l (TMVTobaco mosaic virus), di,... l cc virus c cu trc xon: acid nucleic v protein sp xp dng xon l xo hnh que hay si. Tu loi virus m chiu di, ng knh v chu k lp ca nucleocapsid khc nhau. Virus khm thuc l l virus c cu trc xon c trng, dng hnh que thng, chiu di 300nm, ng knh ngoi 15nm, ng knh li rng 4nm. TMV c cha 5% ARN mch n (ss ARN) v 95% protein. Capsid c cu to bi 2130 capsome hnh chic giy. Mi capsome c cu thnh t 158 gc acid amin, c phn t lng l 17.500 Da. Cc capsome bm vo si ARN xon l xo, c 130 vng xon, vi bc nng vng xon l 2,3nm, v trung bnh c 16,33 capsome gn vo. C mt capsome gn vi 3 nucleotid ca ARN, nn mi vng xon ca ARN c 49 nucleotid. Si ssARN cha 6.390 base nucleotid, c phn t lng l 2 106 D (hnh 10.4).
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2.2.2. Virus i xng khi Cu trc i xng khi c trng cho cc loi virus nh Adenovirus, Reovirus, Herpes virus, Picorna virus,... vi typ i xng 2 mt, 3 mt, 5 mt. Khi kho st cu trc Adenovirus (hnh 10.5), nu quan st thong qua thy gn ging hnh cu, nhng khi quan st k hn nhn thy y l virus khng c mng bao, ng knh khong 70 80nm, c cu trc i xng khi, a din 20 tam gic u, 30 cnh, 12 nh, mi nh l ni gp nhau ca 5 mt. Capsid c cu thnh bi 252 capsome vi phn t lng capsome l 70.000 Da.
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2.2.3. Virus cu trc phc hp Virus cu trc phc hp rt ph bin trong trng hp cc phage, trong c phage T chn ca E. coli. Loi phage ny gm c cc kiu T2, T4 v T6 phn b rt rng ri trong t nhin. y l cc m hnh c nghin cu su, nht l i vi phage T4. Phage T4 c cu to bi 3 b phn: u, c v ui. 2.2.3.1. u Di 95nm, ng knh 65nm, c 20 mt. Capsid c cu to bi 8 loi capsome, ng knh 8nm, tng cng c 212 capsome. Lng protein chim 71 86% trong phage. Bn trong u c ADN si p (ds ADN), u c ni vi ui qua c. 2.2.3.2. C L a lc gic, ng knh 37,5nm. T a c mc ra 6 tua c. 2.2.3.3. ui Gm c bao ui, ng ui, a gc, 6 mu ghim v 6 si ui. Bao ui di 95nm, c 24 vng xon, cu to bi 144 capsome (mi capsome c phn t lng l 55.000Da), u bao ui gn vi vng c. Lng trong bao ui l ng ui vi chiu di v vng xon tng t, ng knh 8nm, rng gia c ng knh 2,5 3,5nm, l ng dn ADN t trong u phage xm nhim vo t bo mn cm. a gc gn vi u cui ng ui cng l a lc gic rng gia c ng knh 30,5nm, trn mc ra 6 si ui v 6 mu ghim.
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Mu ghim di 24nm c chc nng hp ph. Si ui di 140nm c th gp li chnh gia, ng knh 2nm. Si ui c cu to bi 2 loi protein kh ln v 4 loi protein kh nh, c tc dng hp ph vo vng mn cm ca t bo ch (hnh 10.6).
Hnh 10.6. Cu to ca phage
Khi si ui hp ph vo vng mn cm, a gc b kch thch lm co rt bao ui v lm cho ng ui m mu ghim vo t bo ch. Cc capsome ca bao ui pht sinh chuyn ng chuyn v phc hp lm co chiu di ui li 50%, ging s co rt ca cc protein si c.
2.3. Acid nucleic ca virus

Acid nucleic lu gi thng tin di truyn c th ca mi virus, nn rt a dng v l c s tin cy phn loi virus. Cc loi acid nucleic ca virus c th c phn bit da trn cc c tnh sau: ADN hay ARN? Chui n (sssingle strand) hay chui kp (dsdouble strand)? Dng mch thng hay dng vng? H gen cha 1 thnh phn, hay 2, 3 hoc nhiu thnh phn? Nhng virus mang ADN th ch yu l mch kp, cn cc virus mang ARN ch yu l mch n. Phn loi mt s virus n h trn c s acid nucleic vi cc tiu ch nu nh sau: 2.3.1. Cc virus cha ADN a) Chui n (ss ADN) Mch thng: H Parvoviridae (gy bnh st ban tr em) cha c nghin cu nhiu. Mch vng: Thc khun th X 174, M13, fd, f1 ca E. coli. b) Chui kp (ds ADN) Mch thng: + H Adenoviridae (gy nhiu hi chng lm sng khc nhau): Adeno virus typ 1, 8, 14, 21,... + H Herpesviridae (gy bnh mn rp herpes ngi, gy bnh u g), c v bao quanh capsid: Herpes simplex typ 1, Herpes simplex typ 2, Varicella zoster virus, Cytomengalovirus
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(CMV), EpsteinBarrvirus (EBV) lin quan n cc khi u trn ngi. + H Hepadnaviridae (virus ADN gy vim gan), c v bao ngoi: Herpatitis B virus (HBV), Herpatitis delta virus (HDV). + H Poxviridae (gy bnh u ma, bnh u b,...), c v bao ngoi: Pockenvirus, Vacxiniavirus, Affenpockenvirus. + Phage T, P1, P2, Mu ca E. coli, phage PBSX, SPO1 ca B. subtilis, phage P22 ca Salmonella,... Mch vng: H Papovaviridae (gy khi u trn ngi v ng vt): Papilloma, Polioma, Vacuolating virus. 2.3.2. Cc virus cha ARN a) Chui n H Arenaviridae, hnh cu hoc a hnh, c v bao ngoi, gy st xut huyt v vim mng no v trng (Lymphocytosis choriomeningitis virus LMV), cng c loi gy st Lassa. H Bunyaviridae (c 150 typ huyt thanh), capsid cu trc xon, c v bao ngoi, bao gm Hantan virus gy bnh st xut huyt khng do cn trng t v gy bnh thn. H Coronaviridae, cu trc xon, c v bao, c kh nng gy vim ng h hp cp, trong c SARS. H Filoviridae, a hnh thi, c v bao. Chi i din l Filovirus cha Marburgvirus v Ebolavirus gy bnh. H Flaviviridae, gy bnh ly lan qua cn trng tit tc, c v bao ngoi. Cc virus gy bnh chnh bao gm: virus Dengue, virus vim no (encephalitis) Nht Bn B, virus st vng, FSME virus (vim no ma xun), virus vim gan C. H Orthomyxoviridae (gy bnh cm), ARN si (), c v bao envelop, c cc virus gy bnh cm A, B, C cho ngi v ng vt. H Paramyxoviridae, capsid cu trc xon, c v bao, c cc virus gy bnh cm, si, quai b, hp bo h hp (RSV = Respiratory Syncitial virus), bnh Newcastle, Rendepest. H Picornaviridae, l nhm virus nh, gy bnh ch yu ng tiu ho. Cc virus chnh: Poliovirus, Coxsackievirus, echo virus, Hepatitis A virus, Rhinovirus, ngoi ra cn c hng trm virus khc. H Rhabdoviridae, cu trc xon, c v bao ngoi, ch yu gy bnh di. H Retroviridae, hnh cu, c v bao ngoi, c enzym phin m ngc. C nhiu virus gy bnh, c bit c virus gy ung th, v gy bnh AIDS. H Togaviridae, c v bao ngoi: Thnh vin gy bnh chnh l Rubella virus. Cc loi phage cha ARN nh thc khun th MS2, A, f2, R17 ca E. coli,... b) Chui kp (ds ARN) Mch thng: + H Reoviridae: Gy bnh a chy i vi tr em ang b, v nhim khun h hp do Rotavirus.
3. S NHN LN CA VIRUS
Khi nhim sinh vo t bo sng cm th, qua iu khin b my trao i cht cng nh hot ng trao i cht ca t bo m virus tng hp c cc thnh phn cu trc v to ra c cc ht virus mi, pht trin nhn ln. Cc ht virus trng thnh thot ra khi t bo ch bt u chu k nhn ln mi. S nhn ln ca virus mang tnh bng pht chu k, khc hn thi k sinh trng, pht trin 4 pha ca vi khun. Thi gian chu k nhn ln ca cc virus khc nhau l khc nhau. V d: Picornavirus cn 6 n 8 gi, cn Herpesvirus cn hn 40 gi. S lng cc virus "con" (th h sau) c to ra cng khng ging nhau: Picornavirus to ra khong 100.000 ht virus mi trong mi t bo ch, trong khi Poxvirus ch to ra khong 1.000 ht virus mi.
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Quan im chung ca cc nh khoa hc l qu trnh nhn ln ca virus gm 5 giai on: Hp ph Xm nhp Tng hp Lp rp Gii phng.
3.1. S hp ph ca virus ln b mt t bo ch
Cc chuyn ng t nhin to ra va chm ngu nhin gip virus bm vo th th (receptor) c hiu trn t bo cm th. V d: Poliovirus ch hp ph c ln mng t bo ngi v linh trng, m khng hp ph ln t bo ng vt khc v khng c th th. Mt s yu t nh hng n s hp ph ca virus: s lng virus, s hin din ca mt s cation (Ca2+, Mg2+) c tc dng xc tin hp ph, ngc li Al3+, Fe3+ li lm gim hot tnh hp ph ca virus. Nhit khng nh hng n hp ph ca virus.
3.2. Xm nhp ca virus

Ngay sau khi hp ph, acid nucleic virus lp tc xm nhp t bo. Qu trnh xm nhp ny xy ra theo mt trong 3 cch sau: Xm nhp nh c ch ci o: Mng ca virus dung hp vi mng sinh cht ca t bo (v bn cht cc mng ny l ging nhau), v d Paramyxovirus v Retrovirrus, phn nucleocapsid b y vo trong t bo cht. Capsid ca virus b enzym decapsidase ca t bo phn hu, gii phng acid nucleic. Xm nhp nh c ch thc bo hoc nhp bo: Sau khi virus hp ph ln b mt t bo, mng t bo lm vo to thnh khng bo a virus vo t bo cht. Ti y mng khng bo b dung gii, nucleocapsid c gii phng. Bm acid nucleic vo t bo ch: Cc phage sau khi hp ph, si ui co li lm cho mu ui a gc cm vo thnh t bo; cc lysozym u ng ui lm tan mt l thnh t bo, to ra kch thch lm cho capsome ca bao ui co li, ng ui m qua thnh v mng t bo cht, v acid nucleic ca phage c bm vo t bo cht.
3.3. Tng hp cc thnh phn ca virus

Trc tin acid nucleic ca virus cung cp thng tin di truyn cho t bo ch v bt t bo ch tng hp ra cc thnh phn ca virus. Qu trnh tng hp c th c thc hin trong nhn (Adenovirus) hoc trong t bo cht (Poxvirus) ca t bo ch. Giai on ny l giai on n (tim tng) ca virus, c th ko di vi gi n vi ngy. 3.3.1. Virus mang ADN i din l cc Adenovirus, Papovavirus v Herpesvirus. ADN ca cc virus ny c sao chp nhn ln trong nhn ca t bo ch sinh ra mARN. Chng trnh sao chp cn t nht 2 chu k i vi Papovavirus v 3 chu k vi Herpesvirus v Adenovirus. Trong mi trng hp, cc polypeptid ca cc virion c to ra t mARN (hnh 10.7).
ADN virus mARN Protein sm
mARN
Protein trung gian ,
ADN con
mARN
Protein mun
Virus con
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Hnh 10.7. S nhn ln ca Adenovirus v Herpesvirrus
Trong c hai trng hp, genom ca virus c sao chp trong 3 chu k bi ADN polymerase. Trc tin, cc ADN mi c to ra (nh ADN pol t bo ch) c phin m thnh mARN (mARN E1, mARN a) m c dch m thnh protein iu ho (protein rt sm). Protein ny h tr phin m ADN mi thnh mARN E2 (hay mARN b) m c dch m thnh ADN polymerase (protein sm sau) mi tng hp. ADN polymerase ny sao chp genom virus thnh rt nhiu genom virus con m mt phn trong s c phin m thnh mARN L1 (mARN g) m c dch m thnh protein cu trc. Protein cu trc lp ghp vi genom virus con to thnh virion hon chnh. Papovavirus, 2 chu k u c gp lm mt. 3.3.2. Virus mang ARN Cc virus mang ARN c chia lm 3 nhm: 3.3.2.1. Nhm 1: i din l Picornavirus. H gen nhm ny c 2 chc nng, va l mARN gi l si dng (+) (nu phn t ARN c trnh t nucleotid trng vi trnh t ca mARN th c quy c l si (+), cn ngc li l si ()) v cng l khun tng hp si b sung gi l si (). Sau khi xm nhp vo t bo ARN virus gn vo ribosom v c dch m. Sn phm l polyprotein v sau c tch thnh cc polypeptid. Vi chc nng th hai, ARN ca virus c dng lm khun tng hp nn si m vi s tham gia ca ARN polymerase c to ra nh vic tch polyprotein ni trn. Sau si m li c dng lm khun tng hp si dng mi. Si dng ny c lp ghp li to ra virus con (hnh 10.8). ARNvirus si (+) Protein c m ho trong h gen
ARNvirus si ( )
ARN con si (+) Virus con

Hnh 10.8. S nhn ln ca Picornavirus
3.3.2.2. Nhm 2: Gm cc virus m ARN ng vai tr l si (), i din l Myxoviridae, Paramyxoviridae. c im ca nhm ny l ss ARN ch c mt chc nng l lm khun cho qu trnh sao chp, v vy, chng phi c sao chp to ra mARN m t bo li thiu cc enzym thch hp. Ton b si m ca virus cng vi enzym sao chp c trong virion. Qu trnh sao chp tri qua 2 giai on. Giai on u tin to ra mARN m mi ARN ny ch m ho mt chui polypeptid, n giai on th hai s hnh thnh chui y v si dng, si ny c dng lm khun tng hp si m con (hnh 10.9).
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ARN virus si ( )
ARN pol (virus)
mARN
Protein
ARN Pol ph thuc ARN mi tng hp
ARN virus si (+)
ARN virus si ( ) con
Cc virus con trng thnh

Hnh 10.9. S nhn ln ca Orthomyxovirus
3.3.2.3. Nhm 3: i din l nhm Retrovirus, c tnh ca nhm ny l nhim sc th ARN virus sao chp ngc thnh ADN trung gian (hnh 10.10).
ARN virus si (+) ARN mi v Reverse transcriptase ADN ADN trung gian (ci xen vo NST)
ARN (+) mARN (Genome virus con)
Protein
Virus con
Hnh 10.10. S nhn ln ca Retrovirus
3.4. Giai on lp rp
Khi cc phn t protein capsid v acid nucleic c tng hp vi mt lng va (khong 2 3 gi sau khi xm nhim) th giai on lp rp bt u. Cc protein ca capsid bao bc mt phn t acid nucleic to thnh mt ht virus hon chnh. Qu trnh ny c th l t pht, khng cn nng lng, nhng c th b ngn cn bi mt s cht c kh nng gn vo acid nucleic ca virus hoc lm hng cu trc protein. Kt qu l qu trnh lp rp c th to thnh ht virus khng hon chnh: ch c capsid m khng c li acid nucleic. Nhng ht virus ny khng c kh nng gy nhim, tuy nhin, c mang tnh khng nguyn c hiu. Picornavirus, Poxvirus v Reovirus lp rp trong bo tng, cn Adenovirus, Herpesvirus v Papovavirus lp rp trong nhn t bo ch. 3.5. Giai on gii phng Cc ht virus con c gii phng khi t bo sau vi gi n sau vi ngy theo hai cch. Ny chi: Mt s virus c mng bao c gii phng bng cch ny chi qua cc v tr c bit ca mng t bo ch do virus c m. Nh vy, virus s nhn ly mt phn mng t bo ch vi c cu protein c trng ring m virus gi vo y. S ny chi c th xy ra dc mng t bo cht: mng nh ra pha ngoi cng vi nucleocapsid nh ny chi, v cc enzym c bit trn b mt ct
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cho chi ri ra. V d, mng ngoi ca virus Paramyxoviridae c gai glycoprotein (hnh 10.11). Ph hu mng t bo ch: Hng lot virus c gii phng trong mt thi gian ngn bng cch ph v mng t bo ch (phage v mt s loi virus khc) qua ly gii v t thot ra ngoi.
4. S NHN LN CA PHAGE
C 2 loi phage chnh l phage c v phage n ho (tim tng). Phage c: l nhng phage ph hu, gy ly gii t bo ch. Sau khi xm nhp t bo vi khun, chng nhn ln v ph v t bo ch gii phng cc phage mi ra ngoi. y l qu trnh ly gii (lysis). Phage n ho cn gi l phage tim tng hay protophage. Khi phage n ho xm nhp vo t bo vi khun, acid nucleic ca virus gn vo ADN ca vi khun, v tn ti, phn chia nhn ln cng h gen ca vi khun qua nhiu th h m khng ly gii t bo. l qu trnh tim tan (lysogeny). Khi gp iu kin thch hp, acid nucleic ca phage c hot ha, ch huy qu trnh nhn ln v to ra cc phage mi lm tn hi t bo. Qu trnh ny ch din ra vi cc phage mang ds ADN. Trong iu kin thch hp, phage n ho c th tr thnh phage c v gy ly gii t bo ch. Nhng t bo mang phage tim tan gi l t bo tim tan hay t bo sinh dung gii. Qu trnh nhn ln ca phage cng tun theo quy lut chung ca virus (hnh 10.12).
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5. HU QU CA QU TRNH NHN LN CA VIRUS

S k sinh, nhn ln ca virus trong t bo gy nn nhng hu qu nghim trng tip theo i vi t bo ch cng nh vt ch: Ph hu t bo (Cytopathic Effect, CPE): Trong phng th nghim, nh gi mc ph hu t bo ca virus ta xc nh liu hu hoi 50% s t bo (CPD 50 Cytopathic Dose 50%) v n v to plaque (vng tan t bo) (plaque forming unit PFU). Lm sai lch nhim sc th ca t bo: Nhim sc th ca t bo ch c th b gy, phn mnh hoc sp xp li. C th xut hin nhng nhim sc th bt thng v thay i s lng nhim sc th ca t bo. Sinh khi u: Cc nh nghin cu gy ung th thc nghim ng vt do virus, cn ngi, mt s c ch gy ung th c xc nh hon ton. C ch gy khi u c th do virus gy thay i khng nguyn b mt ca t bo, lm mt kh nng c ch do tip xc khi t bo sinh sn. Sinh thai nhi bt thng: Trong 3 thng u ca thi k thai nghn, nu ph n mang thai b nhim virus c th gy nhng hu qu bt thng cho thai nhi (hnh 10.13). To ra ht virus khng hon chnh (defective interfering particleDIP): l nhng ht virus khng c acid nucleic, ch c v protein. DIP khng nhn ln c trong t bo, m ch mang khng nguyn v ca virus hon chnh. To ra cc tiu th: Cc tiu th ni bo c to ra t cc thnh phn ca ht virus, nhng ln hn ht virus, c th trong nhn hoc trong bo tng ca t bo.
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Virus
Ph n c thai cm th
Nhim trng i
Nhim trng m
Nhim trng nhau thai
Nhim trng bo thai
Thai bnh thng
Thai nhim trng (c th b sy thai)
Thai cht
Thai d tt
Hnh 10.13. nh hng ca virus i vi ph n c thai
Chuyn th t bo (Transformation): Genom ca virus tch hp vo genom t bo lm cho t bo th hin tnh trng mi. V d, phage E15 tch hp vo nhim sc th ca Salmonella. Sn xut interferon: Virus ng vai tr cht cm ng kch thch t bo sinh interferon. Cht ny ng vai tr cht cm ng kch thch t bo sinh ra protein khng virus. Bin t bo thnh t bo tim tan (t bo c tim nng sinh ly gii): Genom ca virus (phage) tch hp vo nhim sc th ca t bo (vi khun) tn ti v nhn ln cng genom t bo. Khi c iu kin thch hp, genom ca virus s c hot ho, virus nhn ln v ph hu t bo. Nhng t bo c genom virus tch hp trong nhim sc th c gi l cc t bo tim tan.
CU HI LNG GI
1. Trnh by 3 loi cu trc chnh ca virus. 2. Trnh by phn loi virus da trn acid nucleic virus. 3. Trnh by 5 giai on nhn ln ca virus. 4. Trnh by qu trnh tng hp cc thnh phn ca virus mang ADN. 5. Trnh by cc qu trnh nhn ln ca virus mang ARN. 6. So snh qu trnh nhn ln ca virus mang ARN si () v ARN si (+). 7. Trnh by qu trnh nhn ln v gii phng ca phage. 8. Trnh by hu qu ca qu trnh nhn ln ca virus trong t bo v vt ch.
Chng 11
CC VIRUS GY BNH THNG GP
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1. MYXOVIRUS
Myxovirus c chia lm hai nhm chnh: Orthomyxovirus v Paramyxovirus. Orthomyxovirus gm cc thnh vin l: Cm A, B, C; Paramyxovirus gm cc thnh vin l: quai b, si, cm, virus hp bo ng h hp v mt s virus gy bnh cho ng vt.
1.1. Virus cm (Influenza virus)

Virus cm l thnh vin chnh ca nhm Orthomyxovirus v l cn nguyn ca bnh cm l bnh nhim trng ng h hp cp tnh d thnh dch. Orthmyxovirus gm c 3 typ min dch: cm A, B, C. 1.1.1. c im sinh hc 1.1.1.1. Hnh th v cu trc Virus cm c hnh cu, ng knh 80 120nm. V bao ngoi ca virus l lp lipid kp, trn b mt c cc gai glycoprotein. Vt liu di truyn l ARN chui n (gm c 8 on vi cm A, B v 7 on vi cm C). Protein capsid cng vi ARN to nn nucleocapsid i xng xon. 1.1.1.2. Sc khng Virus cm d b tiu dit bi cc yu t vt l, ho hc nh: tia cc tm nh sng mt tri v cc dung mi ho tan lipid (ete, formalin). 1.1.1.3. Nui cy Virus cm th vi cc t bo tin pht nh: t bo x non bo thai g, bo thai ngi. T bo thng trc cm th l: t bo BHK, vero.
Hnh 11.1. Influenza virus
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1.1.1.4. S nhn ln Mt chu k nhn ln ca virus cn 12 gi. S tng hp ARN ca virus thc hin trong nhn t bo cm th, cn qu trnh tng hp cc protein cu trc v cc thnh phn khc ca virus li xy ra bo tng. Cui cng, s lp rp ca virus thc hin bo tng v chng thot ra khi t bo ch bng hnh thc ny chi c mang theo mng ca t bo ch lm thnh v envelope ca ht virus. 1.1.1.5. Khng nguyn Khng nguyn kt hp b th v khng nguyn ho tan bn cht l nucleoprotein, y l khng nguyn c hiu typ. Khng nguyn ngng kt hng cu, gm c: + Khng nguyn H (Hemagglutinin): c trng cho typ virus. Hin nay c 16 cu trc khng nguyn H t H1 n H16. + Khng nguyn N (Neuraminidase): c trng cho th typ, gm c 9 cu trc khng nguyn t N1 n N9. Khng nguyn H c vai tr quan trng trong kh nng gy nhim ca virus, cn khng nguyn N gip cho virus lan trn trong c th v thc y s phng thch cc ht virus ra khi t bo ch. Cu trc H v N thng d thay i tr thnh typ v th typ mi. 1.1.1.6. Cch gi tn Trc ht gi tn typ virus, sau l ng vt cm th, a danh phn lp, s bnh phm phn lp, nm phn lp, v cu trc H v N. V d: A/ BangKok/3/79/H3N2. B/ Singapor/7/79/H1N2. 1.1.2. Kh nng gy bnh Bnh cm l mt bnh nhim trng cp tnh, gy dch. Virus xm nhp vo c th bng ng h hp, sau thi gian bnh t 1 n 3 ngy th xut hin cc du hiu khi pht nh: st cao 39 40OC, nhc u, au mnh my, mt mi, gai rt v ht hi, chy nc mt, nc mi. Sang giai on ton pht, bnh nhn tip tc st cao 39 40OC, mt mi, au u d di, thng au trn v hc mt, mt sung huyt ngu v c th thy nt xut huyt trn da. mt s bnh nhn c bi nhim phi, bnh nng ln rt nhiu. Din bin ca bnh trong vng 7 ngy th bnh nhn khi (nu khng c bi nhim). Tuy nhin, qu trnh li sc ko di khong 3 6 thng vi cc du hiu n km, mt ng v gim sc lao ng. 1.1.3. Dch t hc Cc t cm thng xut hin vo ma ng xun. S lan trn v trm trng ca t bnh thay i rt nhiu. Cc v dch ton th hay i dch xut hin thng l qung 10 n 15 nm mt ln. Trong , dch cm lan trn v nghim trng nht l do virus cm A gy nn. Trong cc nm 1889 1890, 1918 1919, 1957, 1968 c nhng i dch do cm A. Virus cm B v virus cm C thng gy cc v dch nh, t lan rng. 1.1.4. Chn on vi sinh hc 1.1.4.1. Phn lp v xc nh virus Bnh phm l dch mi hng ly vo cc ngy u ca bnh. Nui cy v phn lp trn cc dng t bo nhy cm. Xc nh virus bng phn ng ngng kt hng cu, sau nh typ bng phn ng trung ho trong t bo hoc c ch ngng kt vi hng cu vi khng th mu. 1.1.4.2. Chn on huyt thanh
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Bnh phm l huyt thanh bnh nhn, ln 1 ly vo tun l u v ln 2 ly cch ln 1 khong 7 10 ngy. Cc phn ng dng xc nh l kt hp b th, ngn ngng kt hng cu. Khng th ln 2 phi tng gp 4 ln so vi ln 1 mi kt lun c bnh nhn c mc bnh. 1.1.5. Phng bnh v iu tr Phng bnh: + Phng bnh khng c hiu nh: Cch ly bnh nhn, nh thuc st khun ng mi hng, st khun dng dng c ca bnh nhn. + Phng bnh c hiu: Hin nay thng dng vaccin bt hot cho nhng ngi cha c min dch, nhng khng th hnh thnh ch khng li virus vaccin m khng c min dch cho vi th typ mi. iu tr: Ch yu l iu tr triu chng, chm sc iu dng v chng bi nhim. Cc thuc khng virus cng c dng l Amantadine v Rimantadin i vi cm A.
1.2. Virus quai b (Mump virus)

Virus quai b l thnh vin ca nhm Paramyxovirus c Johnson v Goodpasture phn lp t nc bt nm 1934. 1.2.1. c im sinh hc 1.2.1.1. Hnh th v cu trc Virus c hnh cu v ng knh khong 120 200nm. V envelope c cu trc lipid kp. B mt ca virus cha cc gai l hemagglutinin (H), neuraminidase (N) v hemolysin (Fglycoprotein). Vt liu di truyn l ARN si n. 1.2.1.2. Sc khng Virus quai b d dng b tiu dit bi cc nhn l ho hc nh: 56OC trong 20 pht, tia cc tm v cc dung mi ho tan lipid. 1.2.1.3. Nui cy Virus c nui cy trn t bo thai g v t bo thng trc vero. 1.2.1.4. S nhn ln Qu trnh tng hp ARN xy ra trong nhn ca t bo. Protein cu trc v cc thnh phn khc ca virus c tng hp ti bo tng. Qu trnh lp rp cc ht virus c thc hin bo tng v gii phng theo phng thc ny chi c ly mt phn mng t bo ch lm v bc. 1.2.1.5. Khng nguyn Khng nguyn kt hp b th v khng nguyn ho tan l thnh phn protein ca nucleocapid. Khng nguyn ngng kt hng cu: Hemagglutinin v Neuraminidase. Khng nguyn ny b c ch bi huyt thanh c hiu khng quai b. 1.2.2. Kh nng gy bnh Bnh quai b l mt bnh truyn nhim cp tnh, gy dch. Virus xm nhp vo c th qua ng h hp, thi k bnh 12 25 ngy, virus nhn ln nim mc ming, kt mc mt, t vo mu v i n cc c quan khc nh mng no, tinh hon, bung trng, tu,... Cc biu hin ca bnh nh sau: Vim tuyn nc bt mang tai: l biu hin lm sng thng gp nht, bnh nhn thy au vng mang tai, kh h ming, st, km n, nhc u, au nhc c khp. Sng tuyn nc bt, thng mt bn, sau 1 2 ngy th sng nt pha bn kia. Tu theo mc sng m c th thy bin i khun mt.
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Vim tinh hon: Thng gp tui thanh, thiu nin, chim 20 30% trng hp, xut hin sau sng tuyn nc bt 7 10 ngy. Bnh nhn thy st cao v rt run v thy sng au tinh hon. Vim bung trng: Gp khong 7% sau tui dy th. Bnh nhn thng thy st v au 1 bn hoc 2 bn h chu. Vim mng no gp 10 35% cc trng hp, nht l tr nh. Ngoi ra, virus quai b cn gy vim tu cp, vim no, vim c tim... nhng cc th ny t gp. Min dch sau bnh kh bn vng, t l ti nhim gp di 3%. 1.2.3. Dch t hc Ngi l tc ch t nhin c nht ca virus quai b. Bnh c trn ton th gii. Bnh thng xut hin trong ma ng xun, c bit l thng 4 hoc thng 5 v ly truyn qua ng h hp, rt d pht trin thnh dch cc a phng hoc cng ng. Quai b him gp tr em di 2 tui v ngi gi nhng hay gp tr t 3 14 tui, thanh nin t 17 20 tui, nam nhiu hn n. Khong 30 40% trng hp nhim quai b m khng c triu chng v y l ngun ly kh trnh nht. 1.2.4. Chn on vi sinh hc 1.2.4.1. Phn lp v xc nh virus Bnh phm ly t nc bt, nc tiu hoc dch no tu. Phn lp trong bo thai g hoc cc t bo nui: T bo s non bo thai g, t bo thng trc vero. Ngoi ra, c th dng k thut min dch hunh quang trc tip pht hin khng nguyn virus trong bnh phm. 1.2.4.2. Chn on huyt thanh Pht hin khng th chng quai b trong huyt thanh bng phn ng kt hp b th, c ch ngng kt hng cu. Nhng tt nht vn l ELISA. 1.2.5. Phng bnh v iu tr Phng bnh: + Phng bnh khng c hiu: Cch ly v trnh tip xc vi bnh nhn. +Phng bnh c hiu: Cng vaccin rt c hiu qu, thng tim chng cho tr sau 1 tui, khi m khng th do m truyn ht hiu lc. iu tr: khng c thuc c hiu chng virus quai b. Do vy, khi mc quai b thng quan tm n chm sc iu dng v iu tr phng nga cc bin chng.
1.3. Virus si (Measle virus)

Nm 1757, Francis Home chng minh virus si l tc nhn gy bnh si. Nm 1954, Enders v Peebles phn lp c virus. Virus si thuc h Paramyxovirus, ging Morbillivirus, loi measle. 1.3.1. c im sinh hc 1.3.1.1. Hnh th v cu trc Virus hnh cu ng knh 120 250nm; v bao ngoi l lp lipid kp, trn b mt c cc gai glycoprotein mang tnh khng nguyn; v capsid i xng xon; vt liu di truyn l ARN si n. 1.3.1.2. Sc khng Virus d b tiu dit bi cc nhn l ho nh: 56OC sau 30 pht, tia cc tm v cc dung mi ho tan lipid (ete, formol). 1.3.1.3. Khng nguyn Virus si ch c 1 typ khng nguyn duy nht v n nh, do vy, nhim si t nhin c min
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dch sut i. 1.3.1.4. Nui cy Virus si c th c nui cy trn t bo mt lp ca ngi hoc kh, cc dng t bo thng trc nh vero, Hep2. 1.3.2. Kh nng gy bnh Virus si ch gy bnh cho ngi. Virus si xm nhp vo c th qua ng mi hng v ng mt. Thi gian bnh t 10 12 ngy. Thi k ton pht vi cc du hiu vim long mt: Chy nhiu nc mt, mi mt sng, kt mc mt v vim long ng h hp trn: Chy nc mi, ht hi, ho, km theo st cao, mt mi. Sau xut hin nt Koplix trong nim mc ming. y l giai on d ly nht. Bnh si in hnh th hin bng pht ban theo th t t trn xung di, sau 5 7 ngy, ban bay dn theo th t khi mc, li nhng vt en trn da, gi l vn da h. Sau mc si, bnh nhn c min dch bn vng sut i. Bnh si gy nhiu tai bin: + Vim phi: Do bi nhim vi khun, rt nguy him vi tr nh. + Vim tai gia. + Vim no cp: T l mc 0,005 0,1% trong cc trng hp b si. + Vim x chai no bn cp: L bnh mn tnh do nhim virus chm no, c th xut hin sau si t 7 10 nm. Ngoi ra, khi mc si, sc khng ca tr em b suy gim tm thi nn c th mc nhiu bnh nhim trng c hi nh: tiu chy, vim gic mc d dn ti m lo,... 1.3.3. Dch t hc bnh si Bnh ly theo ng h hp rt d thnh dch, thng gp vo ma ng xun. i tng cm nhim l nhng ngi cha c min dch bnh mi la tui, nhng gp nhiu tr em tui mu gio v tiu hc. 1.3.4. Chn on vi sinh hc 1.3.4.1. Phn lp virus v xc nh virus Bnh phm: Dch mi hng hoc kt mc. Phn lp trn dng t bo nhy cm B95a. 1.3.4.2. Pht hin khng nguyn virus Bng phng php min dch hunh quang. 1.3.4.3. Chn on huyt thanh Bnh phm: huyt thanh bnh nhn Pht hin khng th bng k thut ELISA, kt hp b th, trung ho,... 1.3.5. Phng bnh v iu tr Phng khng c hiu: Pht hin sm, cch ly bnh nhn, x l cht thi. Phng c hiu: Vaccin si sng gim c c tim cho tr em t 9 11 thng trong chng trnh tim chng m rng. iu tr: Cha c thuc c hiu, vic iu tr nhm gii quyt nhng triu chng bt li, chng bi nhim, nng cao th trng gip bnh nhn t hi phc.
1.4. Virus hp bo h hp (Respiratory Syncytial Virus RSV)
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RSV thuc h Paramyxovirus, c phn lp nm 1956, l cn nguyn chnh gy vim nhim h hp cp tr nh. 1.4.1. c im sinh hc Virus hnh cu, ng knh 150 300nm, ngoi cng l lp v envelope c gai bm bn cht l glycoprotein, tip n nucleocapsid, vt liu di truyn l ARN si n. 1.4.2. Kh nng gy bnh Thi k tr bt u cm nhim virus l t 6 tun 6 thng tui. Du hiu ban u l vim ng h hp trn: st, ho, s mi,... Khong 25 40% trng hp chuyn sang vim ng h hp di vi biu hin triu chng: vim ph qun, vim tiu ph qun, ph qun ph vim. Thng dn n bi nhim vi khun. Sau nhim virus c khng th min dch nhng tn ti khng lu, do tr thng b ti nhim. Tr ln v ngi ln khi nhim virus thng c bnh cnh ca cm lnh, triu chng khng r rng, rt kh chn on lm sng. Dch t: RSV ly lan bng ng h hp, sau nhn ln ti nim mc ng h hp. Bnh thng gp vo ma ng xun, c th thnh dch. 1.4.3. Chn on vi sinh hc 1.4.3.1. Phn lp virus v xc nh virus Bnh phm: Dch mi hng. Nui cy trn dng t bo cm th thch hp. Xc nh virus bng k thut min dch hunh quang. 1.4.3.2. Chn on huyt thanh Bnh phm: Huyt thanh bnh nhn. Tm khng th khng virus bng cc k thut min dch. 1.4.4. Phng bnh v iu tr Cha c vaccin c hiu. Phng bnh bng cc bin php khng c hiu nh v sinh mi hng, pht hin v cch ly bnh nhn. iu tr: Cha c thuc c hiu, iu tr triu chng, chng bi nhim, nng cao th trng.
1.5. Virus cm (Parainfluenzae virus)

1.5.1. c im sinh hc cm l virus a hnh thi, kch thc t 150 200nm, v bao ngoi c cu trc hemaglutinin v neuraminidase gip cho chc nng bm ca virus vo t bo cm th, c kh nng gy ngng kt hng cu ng vt, ng thi l nhng khng nguyn c hiu kch thch c th sinh khng th khng virus, vic pht hin khng th ny trong huyt thanh c gi tr chn on. Vt liu di truyn ca virus l ARN mt si. Virus cm c 4 typ huyt thanh 1, 2, 3, 4. Trong , cm typ 1 cn gi l cm D hay cm Sendai.
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1.5.2. Kh nng gy bnh Bnh thng gp tr em vi biu hin; st, nhim trng ng h hp, i khi bnh cnh ging ho g, cn gi l gi ho g. Bin chng thng gp l bi nhim vi khun: ph cu, lin cu, Haemophilus influenzae, vim tai gia. Khi b bi nhim, tnh trng thng nng ln c th dn n t vong. Bnh ly trc tip qua ng h hp, c th thnh dch trong cng ng. 1.5.3. Chn on vi sinh hc 1.5.3.1. Chn on trc tip Bnh phm: Dch mi hng. Nui cy trn dng t bo cm th thch hp. Xc nh virus bng k thut min dch hunh quang. 1.5.3.2. Chn on huyt thanh Bnh phm: Huyt thanh bnh nhn. Tm khng th khng virus bng cc k thut min dch. 1.5.4. Phng bnh v iu tr Cha c vaccin c hiu. Phng bnh bng cc bin php khng c hiu nh v sinh mi hng, pht hin v cch ly bnh nhn, x l cht thi. iu tr triu chng, chng bi nhim, nng cao th trng.
2. ARBOVIRUS
Arbovirus l nhng virus c th nhn ln c trong cc t chc ca cc ng vt c xng sng (ngi, ng vt c v, chim), cng nh mt s ng vt khng xng sng (ve, mui, dn). Cc virus ny mun truyn bnh t ng vt c xng sng ny ti ng vt c xng sng khc phi thng qua mi gii trung gian l cn trng tit tc.
2.1. Virus vim no Nht Bn (Japanese Encephalitis Virus)

c gi l virus vim no Nht Bn bi virus ny c Hayshi pht hin nm 1934 ti Nht Bn. Ngoi ra, do virus thuc nhm B (Flavivirus) ca Arbovirus cho nn c gi l virus vim no Nht Bn B. 2.1.1. c im sinh hc 2.1.1.1. Hnh th v cu trc Virus c hnh cu, ng knh khong 40 50nm, v bao ngoi l mng lipid kp, capsid i xng
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hnh khi, vt liu di truyn l ARN. Trn b mt ht virus c nhng gai glycoprotein mang tnh khng nguyn. 2.1.1.2. Sc khng Virus b tiu dit bi cc dung mi ho tan lipid (ete, x phng, formalin), tia cc tm, 56OC sau 30 pht. 2.1.1.3. Nui cy Pht trin tt trn t bo mui C6/36. Cng c th nui cy vo no chut nht trng 1 3 ngy tui hoc cy vo lng trng g p 8 9 ngy. 2.1.1.4. Khng nguyn Virus vim no Nht Bn c khng nguyn trung ho, khng nguyn ngng kt hng cu v khng nguyn kt hp b th. 2.1.2. Kh nng gy bnh Bnh vim no Nht Bn do mui t truyn virus, ch yu gp tr em di 15 tui, biu hin lm sng thng gp 2 th sau: Th nh: Bnh nhn ch au u, st nh, mt mi trong 2 3 ngy. Th nng: Do c tn thng no, nn t cc triu chng nh t ngt xut hin cc triu chng vim no cp nng nh: au u d di, st cao, co git, ri lon cm gic, ri lon vn ng, ri lon thc nhiu mc . T l t vong khong 10 12%, cc bnh nhn thot khi thi k nng thng c cc di chng v thn kinh v tm thn. 2.1.3. Dch t hc Bnh vim no Nht Bn lu hnh rng Chu , cc v dch thng xy ra vo ma h. Virus c duy tr cc ng vt c xng sng hoang d, mt s loi chim (chim liu iu) v c bit mt s loi gia sc nui nh ln, b, ch, nga. Vector truyn bnh l mui Culex v Aedes, trong , mui Culex tritaeniorhynchus l vector chnh. 2.1.4. Chn on vi sinh hc 2.1.4.1. Phn lp v xc nh virus Bnh phm: Mu v dch no tu ca bnh nhn sau khi pht bnh 1 3 ngy hoc no t thi cht cha qu 6 gi. Ngoi ra, c th ly bnh phm l vector truyn bnh (mui Culex tritaeniorhynchus). Thng dng 2 k thut sau phn lp virus: + K thut phn lp trn chut nht trng 1 3 ngy tui. + K thut phn lp trn t bo mui C6/36. Xc nh virus ny bng 3 k thut: + K thut ngn ngng kt hng cu. + K thut min dch hunh quang. + K thut ELISA. 2.1.4.2. Chn on huyt thanh Bnh phm: mu huyt thanh kp ca bnh nhn. K thut thng dng hin nay l k thut ngn ngng kt hng cu, ELISA tm ng lc khng th. 2.1.5. Phng bnh v iu tr Phng bnh: + Phng bnh khng c hiu: Dit mui truyn bnh, trnh b mui t. + Phng bnh c hiu: Tim phng vaccin cho tr em di 15 tui, nht l nhng vng c
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dch lu hnh. iu tr: Hin nay cha c thuc iu tr c hiu virus, ch yu l iu tr triu chng kt hp vi chm sc iu dng. i vi th nng th phi iu tr kp thi, trnh cc bin chng v phi phc hi chc nng cho bnh nhn giai on lui bnh.
2.2. Virus Dengue

Virus Dengue c Sabin pht hin nm 1944 v c xp vo nhm B (Flavivirus) ca Arbovirus. 2.2.1. c im sinh hc 2.2.1.1. Hnh th v cu trc Virus Dengue c hnh cu vi ng knh khong 35 50nm. Cu trc ca virus gm c: V envelope bn cht l lipoprotein, v capsid i xng hnh khi, vt liu di truyn l ARN mt si dng. 2.2.1.2. Sc khng Virus b tiu dit bi cc dung mi ho tan lipid (ete, formalin), tia cc tm, 600 sau 30 pht. 2.2.1.3. Nui cy Nui cy virus trn cc t bo nui nh: t bo Hela, t bo mui C6/36. Ngoi ra, c th cy truyn vo no chut nht trng mi 1 2 ngy tui hoc cy vo c th mui Aedes aegypty hoc mui Toxorhynchites. 2.2.1.4. Khng nguyn Virus Dengue c khng nguyn kt hp b th, trung ho v ngn ngng kt hng cu. Do cu trc ca cc quyt nh khng nguyn khc nhau cho nn virus Dengue c chia ra lm 4 typ, k hiu l: D1, D2, D3, D4. 2.2.2. Kh nng gy bnh Bnh st xut huyt Dengue l bnh truyn nhim gy dch do mui truyn. Nhim virus c th gy ra cc bnh cnh sau: St Dengue: Nung bnh 3 15 ngy; thi k khi pht, bnh nhn st cao 39 40OC, rt run, au u, au mi c, khp, sung huyt cng mc mt, au nhc nhn cu, pht ban ngoi da kiu si, i khi thy c chm xut huyt di da, s lng tiu cu bnh thng. St trong vng 3 7 ngy, tin lng tt khng xy ra sc. St xut huyt Dengue: Ngoi cc triu chng nh trn, bnh nhn cn c cc du hiu ca hi chng xut huyt di da, nim mc, xut huyt tiu ho, s lng tiu cu gim. Khi b st xut huyt Dengue rt d dn n sc do gim tun hon (do tng tnh thm thnh mch) vi cc du hiu: mt l, tinh thn vt v, mch nhanh nh, chi lnh, huyt p tt, ni vn tm trn da. Nu khng iu tr kp thi, bnh nhn c th t vong.
2.2.3. Dch t hc
cha virus l ngi, kh v mui, ng ly truyn virus l do mui t. V vy, bnh st xut huyt Dengue c mt ni c mui Aedes aegypti lu hnh nh Chu , Chu Phi, Chu M v Chu c. Vit Nam, do c h thng ao h, knh rch phc tp v kh hu nng m, cho nn bnh st xut huyt Dengue c th gp quanh nm, c bit l vo ma ma.
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2.2.4. Chn on vi sinh hc 2.2.4.1. Phn lp v xc nh virus Bnh phm: + Mu bnh nhn lc ang st. Ngoi ra, bnh phm c th l t chc gan, lch, hch lympho,... ly t t thi cht khng qu 6 gi v c bo qun bi glycerol 50%. + Vector: Mui Aedes aegypti. Bnh phm ly xong phi c bo qun lnh, ring mui phi gi cho sng v gi ngay n phng th nghim. Phn lp virus: Hin nay thng dng mt trong ba phng php sau: + Phn lp trn chut bch. + Phn lp trn mui sng. + Phn lp trn t bo mt lp C6/36. Xc nh virus: Sau khi phn lp, xc nh virus bng mt trong cc phng php sau: + Phn ng kt hp b th. + Phn ng trung ho. + K thut min dch hunh quang trc tip. + K thut PCR (Polymerace chain reaction). 2.2.4.2. Chn on huyt thanh Bnh phm: Mu bnh nhn lc vo vin v ln th hai cch 7 ngy. Cc k thut thng dng hin nay: + Phn ng ngn ngng kt hng cu. + Phn ng kt hp b th. + Phn ng trung ho. Da vo cc phn ng huyt thanh, chng ta tm c hiu gi khng th ln 1 v hiu gi khng th ln 2. T , nu hiu gi khng th ln 2 ln hn ln 1 bn ln th c chn on dng tnh. 2.2.5. Phng bnh v iu tr Phng bnh: Hin nay cha c vaccin phng bnh st xut huyt Dengue, do vy, hn ch mc bnh ny th ch yu da vo phng bnh khng c hiu, l: + Hn ch v tiu dit vector truyn bnh. + Hn ch v trnh mui t. iu tr: Hin nay cha c thuc c hiu khng virus. Ch yu l iu tr triu chng v bin chng kt hp vi nng cao th trng.
3. VIRUS BI LIT (POLIOVIRUS)
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Virus bi lit c tm ra nm 1904, y l mt Enterovirus (virus ng rut) thuc h Picornaviridae. Enterovirus gm: Poliovirus, Coxsackievirus, ECHO virus, virus vim gan A.
3.1. c im sinh hc
3.1.1. Hnh th v cu trc Virus bi lit hnh cu, ng knh khong 27 30nm, capsid c cu to bi 32 capsome i xng hnh khi. Vt liu di truyn l ARN mt si. 3.1.2. Sc khng Virus bi lit c sc khng cao, trong nc c th sng c khong 4 thng, trong phn sng c trn 6 thng, khng b bt hot bi dung mi ho tan lipid. Tuy nhin, virus li d b tiu dit nhit 56OC/30 pht, tia cc tm v cc cht st khun nh oxy gi, dung dch iod.
Hnh 11.3. nh knh hin vi in t ca virus bi lit
3.1.3. Nui cy Hai dng t bo thng trc c s dng phn lp virus bi lit: T bo L20B (dng t bo chut c th th c hiu vi virus bi lit). T bo RD (t bo ung th c vn). 3.1.4. Khng nguyn Virus bi lit c 3 typ huyt thanh khc nhau, l typ I, ging in hnh l Bruhilde; typ II, ging in hnh l Lansing; typ III, ging in hnh l Leon.
3.2. Kh nng gy bnh

Bnh bi lit (Poliomyelitis) l bnh nhim trng cp tnh ly theo ng tiu ho, c th thnh dch. Thi k bnh t 4 35 ngy. Biu hin lm sng ca bnh rt a dng, th lit mm cp ch khong 1%, th vim no v khun khong 1%, cn khong 90% l th n. Biu hin ca hi chng lit mm cp l: gim hoc mt vn ng chi, c b lit mm, nho, gim trng lc, tin trin cp tnh trong 3 4 ngy.
3.3. Dch t hc
Virus bi lit phn b rng ri khp th gii, ly truyn qua ng tiu ho. Nc nhim cht thi, thc n b nhim hoc cn trng trung gian (rui, gin) l ngun ly nhim. Bnh thng xy ra ma h v gp tr em di 15 tui. T nm 2000, Vit Nam cng b thanh ton bi lit, tuy nhin bi lit vn lu hnh ti mt s ni trn th gii, do chng ta vn phi
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tip tc gim st cc trng hp lit mm cp theo thng quy ca T chc Y t Th gii.
3.4. Chn on vi sinh

3.4.1. Phn lp v xc nh virus Bnh phm: Phn ly trong vng 0 14 ngy k t khi lit, 2 mu ly cch nhau 24 gi. Phn lp virus trn 2 dng t bo nhy cm: T bo L20B, t bo RD. Xc nh s c mt ca virus l nhng m hoi t t bo. nh typ virus bng phn ng trung ho trn nui cy t bo. 3.4.2. Chn on huyt thanh Bnh phm: Huyt thanh bnh nhn c ly 2 ln cch nhau khong 7 ngy tm ng lc khng th. Cc phn ng thng dng: Phn ng trung ho trn nui cy t bo.

Phng bnh: + Phng bnh khng c hiu: m bo v sinh thc phm, v sinh ngun nc v tng cng dit cn trng gieo mm bnh (rui gin). Ngoi ra, phi pht hin sm bnh nhn cch ly v x l cc cht thi, dng c lin quan n bnh nhn bng chloramin 1% trong 1 gi. + Phng bnh c hiu: Hin nay c 2 loi vaccin c s dng l vaccin bt hot Salk v vaccin sng gim c Sabin. iu tr: Ch yu l iu tr triu chng, phc hi chc nng phng bin chng v nng cao th trng cho bnh nhn.
4. ROTAVIRUS
c Bishop pht hin nm 1973 khi quan st di knh hin vi in t mnh sinh thit nim mc rut mt tr em cht v tiu chy. Sau c t tn l Rotavirus thuc h Reoviridae.
4.1. c im sinh hc
4.1.1. Hnh th v cu trc Di knh hin vi in t, virus trng ging hnh bnh xe, ng knh khong 70 75nm, c capsid i xng hnh khi gm c hai lp bao bc quanh mt ARN si kp. Rotavirus khng c v envelope. 4.1.2. Sc khng Rotavirus c th tn ti nhiu ngy trong phn nhit thng v khng chu tc dng ca cc cht ho tan lipid. Virus c kh nng tn ti lu trong bung bnh, nn t l nhim trng bnh vin kh cao. Rotavirus d b bt hot bi ho cht EDTA v nhit cao hn 45OC. 4.1.3. Nui cy Rotavirus c th c nui cy trn t bo tin pht nh: t bo rut, t bo thai ngi, t bo thai ln.
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4.1.4. S nhn ln trong c th

Rotavirus thng gy nhim cc t bo c nhung mao trng thnh rut non v qu trnh nhn ln ca chng din ra hon ton trong bo tng ca t bo.
4.2. Dch t hc v kh nng gy bnh

4.2.1. Dch t Nhim Rotavirus xut hin khp th gii. n 3 tui, gn nh mi mt c th b nhim Rotavirus mt ln. vng kh hu n ho, nhim Rotavirus thng xut hin vo ma ng lnh. vng nhit i, nhim Rotavirus c xu hng xut hin sut nm nhng nhiu nht l ma thu ng. Rotavirus ch yu gy tiu chy cp tr di 3 tui nhng cng c th gy bnh ngi ln, c bit l nhng i tng suy gim min dch, nhim HIV. S ly truyn ca Rotavirus l theo con ng tiu ho. Vit Nam, t 1980 xc nh Rotavirus l cn nguyn gy vim d dy, rut cp tnh tr em, y l mt bnh rt ph bin ng th hai sau nhim trng h hp cp tnh tr em. 4.2.2. Kh nng gy bnh Rotavirus gy vim d dy, rut cp tnh. bnh thng ch 1 2 ngy. Ton pht xy ra t ngt vi cc triu chng st nh, nn, tiu chy nhiu ln, phn thng c nhy v khng c mu. Du hiu mt nc nh hay nng tu theo mc tiu chy.
4.3. Min dch

Tr nh di 3 thng t mc bnh do khng th c t m. Cc loi khng th: IgA tit trong sa m, chim t l cao v tn ti n 24 thng. IgA tit nim mc rut non, l c s nghin cu vaccin ng ung. IgM xut hin sm trong mu giai on cp tnh. IgG tng cao giai on hi phc.

Bnh phm: L phn ly trong tun u ca bnh. Phn lp virus: t lm. Chn on nhanh pht hin virus t bnh phm bng mt trong k thut sau: + K thut min dch men (ELISA): Hin nay c s dng nhiu nht.
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+ Min dch hunh quang (Immuno Fluorescene). + Min dch phng x (RIA). + Ngng kt hng cu (hoc ht Latex) th ng. + Tm v quan st ln v hnh thi ht virus bng knh hin vi in t.

Phng bnh: Hin nay cha c vaccin phng bnh. Phng bnh khng c hiu vi nhng cng vic sau: v sinh n ung, v sinh mi trng, phng chng nhim trng bnh vin. iu tr: Ch yu l bi ph nc v in gii, kt hp vi nng cao th trng cho bnh nhn n khi hi phc hon ton.
5. CC VIRUS GY VIM GAN (Hepatitis viruses)

Cc virus gy vim gan l nhng virus c i tnh vi t bo gan, nhng chng c cu trc, ng xm nhp, c ch lan truyn khc nhau. Nhng virus ny xm nhp, nhn ln v gy tn thng t bo gan. Hin nay cc virus gy vim gan c chia thnh 5 loi chnh: A, B, C, D, E.
5.1. Virus vim gan A (Hepatitis A virus HAV)

5.1.1. c im sinh hc 5.1.1.1. Hnh th v cu trc HAV l typ th 72 ca Enterovirus, thuc h Picornavirus, khng c v envelope, ngoi cng l v capsid i xng khi bao bc vt liu di truyn l ARN mt si dng.
Hnh 11.5. nh hin vi in t ca virus vim gan A
5.1.1.2. Sc khng HAV khng b bt hot bi cc dung mi ho tan lipid, nhng li d dng b bt hot bi tia cc tm, ho cht st trng, 100OC sau 5 pht. 5.1.1.3. Nui cy C th nui cy HAV trn t bo lng bi phi ngi, t bo vero,... 5.1.1.4. Khng nguyn HAV ch c mt khng nguyn chung l HAAg, do , virus ny ch c 1 typ ng nht. i vi nhng ngi mc bnh vim gan A th c th s p ng min dch vi khng th lp IgG tn ti trong nhiu nm v c th l sut i. 5.1.2. Kh nng gy bnh HAV gy bnh vim gan cho ngi, bnh d ly lan thnh dch. Thi gian bnh khong 30
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40 ngy ri biu hin cc triu chng nh: st nh, mt mi, chn n, nc tiu vng, vng da, nim mc, men gan tng cao, rt him c cc triu chng nng. Khong 40 60% ngi mc HAV khng c biu hin lm sng, y l ngun ly nguy him.
5.1.3. c im dch t
HAV c mt khp th gii v ly truyn qua ng tiu ho. Ngun ly chnh l ngi mang virus khng biu hin triu chng v ngi bnh. i tng nhim HAV ch yu l tr em v ngi sng thiu v sinh. Bnh vim gan A c t l mc cao vng nhit i, c bit l cc nc ngho c iu kin v sinh thp km.
5.1.4. Chn on vi sinh hc

5.1.4.1. Phn lp v xc nh virus Bnh phm l phn hoc mnh sinh thit gan. C th xc nh trc tip virus trong bnh phm bng knh hin vi in t hay k thut min dch hunh quang v min dch phng x. Ngoi ra, c th nui cy phn lp virus trong cc t bo vero hoc t bo lng bi phi ngi. 5.1.4.2. Chn on huyt thanh Bnh phm l huyt thanh bnh nhn. C th tm thy IgM ngay t nhng ngy u ca bnh bng phn ng ELISA, sau l IgG cng tm c bng cc phn ng kt hp b th, phn ng trung ho. 5.1.5. Phng bnh v iu tr Phng bnh: + Phng bnh khng c hiu: V sinh n ung, qun l bnh nhn v x l tt cht thi, dng ca bnh nhn,.... + Phng bnh c hiu: Dng vaccin. iu tr: Ch yu l chm sc iu dng v iu tr triu chng.
5.2. Virus vim gan B (Hepatitis B virus HBV)

5.2.1. c im sinh hc 5.2.1.1. Hnh th v cu trc HBV thuc h Hepadnaviridae. Nm 1970, Dane pht hin thy virus trong huyt thanh bnh nhn bng knh hin vi in t di dng ht hnh cu phc tp, ng knh khong 42nm v vng li m kch thc khong 28nm. Ht ny c gi l ht Dane. Cu trc ht Dane gm: V ngoi dy 7nm cu to bi 3 protein cu trc lm cho virus c hnh cu; capsid i xng hnh khi to thnh li; vt liu di truyn l ADN hai si khng khp kn. 5.2.1.2. Sc khng Virus b bt hot bi tia cc tm, nhit 100OC sau 5 pht 5.2.1.3. Nui cy Hin nay cha tm c h t bo nui cy thch hp cho HBV.
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5.2.1.4. Khng nguyn HBV c 3 loi khng nguyn chnh: + HBsAg l khng nguyn b mt. y l khng nguyn c s thay i gia cc th typ, gm c 4 typ ph: adw, ayw, adr, ayr. + HBcAg l khng nguyn li nm trung tm ca ht virus. Mun pht hin c khng nguyn ny phi ph v ht virus. + HBeAg l khng nguyn c ngun gc t nucleocapsid, thng thay i cc th typ v gm c 2 typ ph: HBeAg/1, HBeAg/2. 5.2.1.5. Khng th Khi c th nhim HBV th s sinh cc khng th tng ng: + Khng th khng HBsAg: Xut hin rt mun sau khi nhim HBV, c tc dng chng virus nn khi xut hin th bnh cnh c ci thin. + Khng th khng HBcAg: C sm giai on bnh, nhng nu ko di th bnh nhn s mc vim gan mn tnh. + Khng th khng HBeAg: xut hin rt mun, thng thi k lui bnh v hi phc. 5.2.2. Kh nng gy bnh HBV ch gy bnh cho ngi, thi gian bnh thng 40 90 ngy hoc di hn. Thi k khi pht v ton pht thng biu hin rm r, cp tnh vi cc triu chng: St cao, mt mi, chn n, mt ng, nc tiu vng, vng da, vng mt. Thng bnh nhn bnh phc sau 4 tun vi triu chng i tiu nhiu, nc tiu trong dn v bnh nhn tr li n kho bnh thng. Tuy nhin, c khong 5 10% vim gan B cp c th tr thnh mn tnh v c cc bin chng x gan hay ung th gan. 5.2.3. c im dch t HBV ly truyn ch yu qua 3 con ng chnh: ng mu, ng tnh dc v t m sang con. Hin nay, c nhiu con ng c th dn n nhim virus ny, v d: tim truyn (ch yu l tim chch ma tu), gi mi dm v cc con ng khc nh ct tc, nh rng, chm cu. Do vy, vim gan B c th gp mi la tui v c t l cao. Ngoi ra, vim gan B cn c bit l cn nguyn ch yu dn ti suy gan v ung th gan. 5.2.4. Chn on vi sinh hc Hin nay, chn on vim gan B thng da vo cc phn ng huyt thanh tm cc marker (du n) ca HBV xc nh virus v cc dng nhim virus. Ngoi vic chn on da vo cc marker ca HBV, trn thc t ngi ta c th dng k thut PCR (polymerase chain reaction) xc nh s c mt ca virus trong mu bnh nhn.
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5.2.5. Phng bnh v iu tr Phng bnh: + Phng bnh khng c hiu: Rt quan trng, l trnh s ly nhim HBV theo nhiu ng khc nhau, cc dng c y t phi m bo nguyn tc v trng. + Phng bnh c hiu: Hin nay c vaccin c sn xut bng huyt tng ca ngi nhim HBV v vaccin ti t hp phng vim gan B. iu tr: Ch yu l iu tr triu chng, ngh ngi, ch dinh dng hp l. Hin nay, ngi ta thng s dng cc thuc khng virus khng c hiu nh: interferon trong iu tr vim gan virus.
5.3. Virus vim gan C (Hepatitis C virus HCV)

Virus gy vim gan C trc y c xp vo nhm virus "non" A, "non" B ("khng" A, "khng"B). n nm 1988, Shikata xp virus "khng" A, "khng" B thnh mt virus ring bit l virus gy vim gan C (HCV). Virus ny thuc h Flaviviridae. 5.3.1. c im sinh hc 5.3.1.1. Hnh th v cu trc HCV c hnh cu, ng knh khong 40 60nm, v envelope c cu trc lipid kp, v capsid i xng khi v trong cng l ARN si n. 5.3.1.2. Sc khng Virus b bt hot bi dung mi ho tan lipid, tia cc tm, nhit cao. 5.3.2. Kh nng gy bnh HCV gy bnh cho ngi, thi gian bnh t 2 tun cho n 3 4 thng. Trong : 95% s ngi c triu chng khng r rng v 5% bnh nhn c ri lon tiu ho, mt mi do cc tn thng t bo gan. Khong 50 70% bnh nhn vim gan C chuyn thnh mn tnh, iu ny rt nguy him, rt d dn n x gan v ung th gan.
Hnh 11.7. nh hin vi in t ca virus vim gan C
5.3.3. c im dch t Bnh ly truyn ch yu bng ng mu, do , i tng thng gp nhng ngi tim chch ma tu hoc mc cc bnh v mu nh hemophylie, leucemie, suy tu xng. 5.3.4. Chn on vi sinh hc Ch yu l da vo tm khng th khng HCV bng k thut ELISA v c th sinh thit t bo tm tn thng bo tng hoc nhn t bo gan. Kt hp vi du hiu tng men gan trong mu.
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5.3.5. Phng bnh v iu tr Phng bnh: + Ch yu l phng bnh khng c hiu nh kim sot mu v cc ch phm mu, tuyn truyn gio dc np sng lnh mnh. + Phng bnh c hiu: Cha c vaccin phng HCV. iu tr: Tng t nh i vi vim gan B.
5.4. Virus vim gan D v virus vim gan E (Hepatitis delta virus HDV, Hepatitis E virus HEV)
5.4.1. Virus gy vim gan D (HDV) Cu trc: L virus khng hon chnh gm mt si ARN, v bao ngoi l HBsAg ca HBV, v trong l khng nguyn delta (AgHD). Dch t v kh nng gy bnh: Vim gan D ly truyn theo ng mu, bnh thng lu hnh nhng vng c t l nhim HBV cao. Ngi ta thy, c th nhim HDV ng thi vi HBV hoc bi nhim HDV ngi mang HBV mn tnh. Chn on: Pht hin khng nguyn delta hoc tm khng th khng delta bng k thut ELISA. 5.4.2. Virus gy vim gan E (HEV) c xc nh vo nm 1985 v xp trong h Caliciviridae. Cu trc: Gm ARN mt si dng xon, v capsid i xng 20 mt, kch thc khong 27nm v khng c envelope. Dch t hc v kh nng gy bnh: HEV ly truyn qua ng tiu ho, c th gy ra thnh dch hoc lu hnh nhng vng v sinh km v c lin quan n ngun nc b nhim. Vit Nam cng xc nh c dch HEV ng bng sng Cu Long. HEV gy bnh thng nh, lnh tnh v khng gy ra cc bin chng nng, nhng cng c th gy vim gan nng cho ph n c thai. Chn on: C th xc nh s c mt ca virus bng knh hin vi in t hoc tm khng th khng HEV.
6. HERPESVIRIDAE
6.1. Cc c im chung h Herpesviridae

6.1.1. Hnh th v cu trc Cc virus thuc h Herpesviridae c cu trc hnh cu vi kch thc khong 180 200nm v gm cc thnh phn sau: Ngoi cng l lp v envelope ly t mng nhn t bo ch. Capsid i xng hnh khi, gm c 162 capsome. Vt liu di truyn l ADN hai si thng. 6.1.2. Sc khng khng rt yu. Nu gy nhim vo m nui cy th cng khng th gi gn c lu hn 2 thng. 6.1.3. Nui cy Virus c th c nhn ln mi trng nui cy l t bo ngi hoc kh. 6.1.4. c im nhn ln
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Virus bm vo receptor ca t bo cm th ri ho nhp v envelope vo mng nguyn tng. Sau , v capsid c ci b trong bo tng v phc hp ADNprotein xm nhp vo trong nhn t bo. Ti nhn, ADN sao m thnh mARN v protein cu trc. Qu trnh lp rp ca cc ht virus c thc hin trong nhn t bo v v envelope c hnh thnh t mng nhn sau khi lp rp xong th cc ht virus gii phng khi t bo. 6.1.5. Phn loi Cc Herpes virus gy nhim cho ngi gm: Herpes simplex, virus gy nhim ng tit niu v mi. Varicella zoster, virus gy thu u v zona. Epstein Barr, virus gy cc bnh: tng bch cu n nhn nhim trng, u t bo lympho B, ung th hu hng. Cytomengalovirus.
6.2. Virus thu u v zona (varicella and zoster virus)
6.2.1. Kh nng gy bnh Virus Varicella zoster gy ra hai thc th lm sng khc nhau, l: Thu u (Varicella) v Zona (Herpes zoster). Thu u: Do nhim virus qua ng h hp, virus c nhn ln v cui cng l nhim virus mu. S xut hin nhim virus mu ny l nguyn nhn ca cc tn thng lan to bnh thu u. Biu hin lm sng l sau khi bnh 10 21 ngy th xut hin st v cc tn thng trn da t dt sn n cc nang nc xut hin thn, mt, ri lan nhanh chng ra cc vng khc ca c th. Cc nang nc ny c th tr thnh mn m do bi nhim lin cu, hay t cu vng. Zona: Biu hin lm sng cng l nhng mn nc, nhng khc vi thu u l zona ch gp ngi ln. Zona l vim thn kinh, do cc mn nc mc dc theo dy thn kinh b vim v kt hp vi s au khng khip ca bnh nhn. Cc mn nc ny cng c th ho m khi c bi nhim. Cng mt loi virus gy ra c hai th bnh v virus zona c coi l s ti hot ca virus thu u tim tng trong cc hch giao cm sau khi b bnh thu u. S ti hot ng ca virus thu u thng gp nhng ngi b suy gim min dch, ri lon min dch t bo. Zona rt hay xy ra nhng ngi mc bnh c tnh, AIDS. 6.2.2. Dch t hc Ngi l bnh duy nht bit ca virus varicella zoster. Virus ny ly truyn qua ng h hp v thng xy ra vi tr em hay ngi ln b suy gim min dch. S ly nhim virus c hai gii v cc c th ca tt c cc chng tc u d dng nh nhau. Virus varicella zoster lu hnh, a phng trong qun th ln, song bnh c th tr thnh dch trong cc c th nhy cm theo ma cui ng, u xun. 6.2.3. Chn on vi sinh hc 6.2.3.1. Chn on trc tip
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Lm tiu bn Tzank: Bng cch pht y tn thng (mn nc) ln lam knh ri nhum bng Giemsa chng minh l cc t bo khng l nhiu nhn. Nhum hunh quang trc tip cc t bo ly t y tn thng da pht hin virus. 6.2.3.2. Chn on huyt thanh Min dch hunh quang l phng php pht hin khng th khng khng nguyn mng ca virus Varicella zoster. Min dch ELISA: Cn gi l min dch gn men v cng dng pht hin khng th khng khng nguyn mng ca virus.
6.2.4. Phng bnh v iu tr

Phng bnh: Cc phng php phng bnh khng c hiu nh: Cch ly bnh nhn hoc trnh tip xc. Hin nay ngi ta dng vaccin gy min dch ch ng cho nhng i tng c nguy c t c kt qu rt tt. iu tr: Ngoi vic iu tr triu chng v chm sc iu dng nhng ch tn thng trnh bi nhim, c th dng cc loi thuc khng virus nh acyclovir, th cc tn thng da s mau lnh hn v gim bin chng.
6.3. Herpes simplex virus (HSV)

Mt s c im sinh hc v kh nng gy bnh HSV c hai typ huyt thanh l HSV1 v HSV2. V tr nhim: + HSV1 gy nhim phn trn lng nh: mm, mi, da. + HSV2 gy nhim phn di lng, c bit l ng sinh dc, tit niu. ng ly nhim: + HSV1 ly theo ng tiu ho, h hp hoc trc tip mi mi. + HSV2 ly nhim theo ng tnh dc. Nhim HSV ngi biu hin thnh 2 giai on: + Nhim ln u (s nhim): Bnh thng nng, biu hin ton thn v c th t vong. Sau gy s nhim, HSV vn tn ti trong c th dng tim n. + Ti nhim HSV c th vi typ huyt thanh khc. Trng hp nhim HSV c khng th trung ho th ch nhim HSV khu tr m khng nhim ton thn.
Hnh 11.9. Herpes simplex virus
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7. VIRUS DI (Rabies virus)

Rabies virus thuc nhm Rhabdovirus.
7.1. c im sinh hc
7.1.1 Hnh th v cu trc Virus di c hnh tr ging nh hnh vin n: mt u trn, u kia dt, chiu di trung bnh t 100 300nm, ng knh 70 80nm. Vt liu di truyn l ARN mt si m, bao quanh c lp v capsid v v envelope. 7.1.2. Sc khng Virus d b bt hot bi nhit 56OC/30 pht hoc 80OC/10 pht, tia cc tm, nh sng mt tri v cc dung mi ho tan lipid nh: ete, formalin, x phng. 7.1.3. Nui cy Virus di c th nui cy vo bo thai g 7 ngy v t bo thng trc nh: t bo vero, t bo thn chut t BHK21.
Hnh 11.10. nh hin vi in t ca virus di
7.1.4. S nhn ln Virus ho v envelope vi mng t bo ch v bm nucleocapsid vo bo tng. Qu trnh sao m, gii m, tng hp protein cu trc, s lp rp xy ra bo tng. Virus c gii phng theo hnh thc ny chi. 7.1.5. Khng nguyn Virus di gm c cc loi khng nguyn sau: Protein G: Khng nguyn kch thch c th sinh khng th trung ho. Protein N: Nm trong phn li ca virus.
7.2. Phn loi

Theo tnh cht sinh hc, chia 2 loi: Virus di hoang di. Virus di c nh: Khi nui cy lu di (thng hn 50 ln cy truyn) virus di s tr thnh
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virus di c nh. Virus ny c thi k bnh ngn (7 ngy), c bo tn bng nui cy trong phng th nghim, khng th tn ti trong iu kin t nhin...
7.3. Kh nng gy bnh

Virus di c kh nng gy bnh cho ngi v cc ng vt mu nng. Virus xm nhp vo c th ngi t ng vt b di qua vt cn, virus ny c i tnh vi t bo thn kinh, sau khi xm nhp, n tn ti v nhn ln ti vt thng, ri lan theo thn kinh ngoi bin n cc dy thn kinh trung ng v no vi cc tn thng c hiu. Vo giai on cui, ton b h thn kinh trung ng cng nh mt s m ngoi tuyn nc bt cng b nhim virus. Bt k ngi hay ng vt, khi m c biu hin bnh ny th tin trin rt nng n v kt thc l cht. Biu hin lm sng ngi: Sau khi bnh 1 3 thng th xut hin tin triu chng ko di 1 4 ngy vi cc triu chng nh st nh, nhc u, bun nn, chy nc mt, nc mi. n thi k ton pht c th gp 2 th bnh l: th co cng v th lit. Bnh nhn cht trong tnh trng suy h hp v ri lon tun hon.
7.4. Dch t
Virus di lu hnh khp th gii, nhng tp trung nhiu cc vng nhit i nh: ng Nam , Chu Phi, Nam M, cha ch yu l ng vt mu nng b di nh: ch, mo,... Virus truyn t ng vt sang ng vt v ngi qua vt cn, vt co,... Vit Nam, bnh di tp trung ch yu ng bng sng Hng v sng Mkng.

Rt t lm v ly bnh phm kh khn v khng c ngha trong iu tr. C cc phng php chn on sau: 7.5.1. Tm tiu th Negri M no ch nghi di nhum tiu bn tm th Negri, l nhng tiu th do virus gy bin i t bo bt mu eosin, kch thc 0,25 25nm. 7.5.2 Phn lp virus Bnh phm l nc di ngi hoc ch lc ang mc bnh, hoc no khi cht. Tim truyn bnh phm vo chut trng mi , sau 3 4 ngy, chut xut hin lit mm, m ly no chut v xc nh virus bng phn ng min dch hunh quang. 7.5.3. Xc nh khng nguyn C th ly nc di hoc no ca bnh nhn hay sc vt b di pht ln tiu bn, khng nguyn s c pht hin bng k thut min dch hunh quang.

Tim phng vaccin di cho ch, mo trong cm dn c hng nm. Ngi b ch, mo nghi di cn th phi tim phng ngay. iu tr d phng trng hp b ch, mo cn: + iu tr ti ch: Ra vt cn bng nc x phng c 20%, sau l nc mui sinh l v st khun bng cn. + iu tr bng vaccin v khng huyt thanh khng di: * Nu vt cn vo ch nguy him (gn u, su) hoc nu ch b mt tch, b cht, hoc ch con th phi tim huyt thanh khng di v tim vaccin ngay. * Nu vt cn nng, xa u, th nht ch li v theo di: Nu sau 10 ngy ch vn sng bnh thng th khng cn tim vaccin. Nu sau 10 ngy b ch m hoc cht th phi tim huyt thanh khng di v vaccin ngay. Vaccin phng di cho ngi hin nay l: vaccin Fuenzalida hoc vaccin verorab.
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8. VIRUS SUY GIM MIN DCH NGI (HIV: Human Immunodeficiency Virus)
8.1. c im sinh hc
8.1.1. Hnh th v cu trc HIV thuc h Retroviridae. Virus c hnh cu, ng knh khong 100nm. Ht virus hon chnh (virion) c cu trc gm 3 lp:
Lp v ngoi l mt mng lipid kp, gn trn mng l cc gai nh, l cc phn t glycoprotein c khi lng phn t 160 kilo dalton (vit tt: gp 160). Gai nh gm 2 phn: + Glycoprotein mng ngoi c khi lng phn t 120 kilodalton (gp 120), y l mt khng nguyn d thay i lm cho khng th khng c kh nng bo v c th v gy kh khn cho vic iu ch vaccin. + Glycoprotein xuyn mng khi lng phn t 41 kilo dalton (gp 41). Lp v trong (v capsid) gm 2 lp: + Lp ngoi hnh cu, cu to bi protein c khi lng phn t 18 kilodalton (p18) vi HIV2 v p17 vi HIV1. + Lp trong hnh tr, cu to bi cc protein c khi lng phn t l 24 kalodalton (p24). y l khng nguyn quan trng chn on nhim HIV. Li: Gm 2 si ARN l b gen di truyn ca HIV, men sao m ngc RT (Reverse Transcriptase) v mt s enzym gip cho qu trnh tng hp virus mi. 8.1.2. Tnh cht nui cy Nui cy trn t bo lympho ngi v t bo thng trc Hela c CD4. 8.1.3. Sc khng HIV d b bt hot bi cc yu t l hc, ho hc. HIV b tiu dit nhit 56OC/30 pht. HIV b bt hot vi mt s ho cht nh oxy gi, Javen,... nhng khng b tiu dit bi tia cc tm.
8.2. S xm nhp vo t bo v nhn ln ca HIV

8.2.1. S hp ph ln b mt t bo
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HIV bm vo b mt t bo cm th nh s ph hp gia cc im tip nhn (receptor) ca t bo ch vi gp 120 ca virus. 8.2.2. S xm nhp vo t bo Sau khi bm c vo cc im tip nhn ca t bo ch, phn t gp 41 ca HIV cm su vo mng t bo to nn s ho nhp ca v HIV vi mng t bo ch. Nh m ARN ca virus chui vo bn trong t bo.
Hnh 11.12. S xm nhp v nhn ln ca HIV 1. Hp ph ln b mt v xm nhp vo t bo. 2. Sao m ngc. 3. Tch hp vo nhim sc th t bo ch. 4. Lp rp cc thnh phn cu trc. 5. Gii phng ht virus mi
8.2.3. S nhn ln trong t bo Sau khi vo trong t bo, nh enzym sao m ngc, ADN ca virus c to thnh t khun mu ARN, sau l qu trnh tch hp ADN virus vo ADN ca t bo ch. Tip , l qu trnh sao m, dch m tng hp cc thnh phn ht virus mi. 8.2.4. Gii phng ht virus mi Cc virus mi c tng hp n gn mng t bo ch, y mng ny ny chi, cc ht virus c gii phng tip tc xm nhp vo t bo khc. 8.2.5. Hu qu ca qu trnh nhim HIV HIV c th xm nhp v nhn ln nhiu loi t bo nh t bo mu v bch huyt, t bo no, t bo d dy, rut, da gy nn cc triu chng bnh l ti y. c bit, HIV xm nhp ch yu vo t bo lympho T CD4 (+) (t bo h tr lympho B trong sn xut khng th) v lympho TC trong min dch t bo). Cc t bo lympho b suy gim ko theo s suy gim ca h thng min dch dn ti bnh nhn b nhiu loi nhim trng c hi nh vim phi, vim da, vim rut, vim hng do vi khun, nm, virus, v ung th, c bit l Sarcom kaposi.
8.3. ng ly truyn
Ly truyn qua ng tnh dc: y l ng ly truyn ph bin, nguy c cao hn nhng ngi ng tnh luyn i nam. Ly truyn theo ng mu: Do truyn mu hoc cc sn phm ca mu. C th ly truyn qua bm kim tim, cc dng c phu thut, chch, xm, rch. c bit t l cao nhng ngi nghin
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chch ma tu. Nguy c ly nhim qua ng mu rt cao, trn 90%. Ly truyn t m sang con: S ly truyn c th xy ra khi c thai, trc, trong v sau khi . Nguy c ly truyn qua ng ny ti 40 50%.
8.4. Min dch

8.4.1. S to thnh khng th Khi HIV xm nhp, c th c kh nng to ra cc khng th sau: Khng th trung ha: Khng th ny c hiu typ chng li cc khng nguyn v (quan trng nht l khng nguyn gp 120). Khng th ny c vai tr bo v v n ngn cn qu trnh xm nhp vo trong t bo. To khng th c st t bo: Cc khng th IgG kt hp c hiu vi cc khng nguyn virus dn ti tan t bo b nhim HIV v gii phng cc ht virus. 8.4.2. Min dch t bo Hnh thnh cc t bo lympho TC (T c). Cc t bo ny kt hp c hiu vi khng nguyn ca HIV (xut hin trn t bo ch) v tiu dit cc t bo ny cng ci cc ht virus trong t bo. 8.4.3. S n trnh h thng min dch ca HIV HIV n trnh h min dch bng hnh thc ch yu l bin d khng nguyn. iu ny gy kh khn cho vic sn xut vaccin phng bnh AIDS, HIV nh vo cc t bo min dch quan trng l TCD4 (+), i thc bo, hay HIV tn ti dng provirus.
8.5. Chn on vi sinh

8.5.1. Pht hin khng th chng HIV Thng c dng nhiu nht trong chn on nhim HIV v nhng k thut pht hin khng th ph hp vi vic xt nghim hng lot mu mu cho kt qu nhanh, nhy cao nhng khng pht hin c giai on "ca s". Hai k thut sau c p dng ph bin: K thut ngng kt Latex nhanh (SERODIA). K thut min dch enzym ELISA. 8.5.2. Pht hin HIV Phn lp HIV: HIV c th phn lp trn t bo lympho hoc t bo Hela c CD4+. Phng php ny rt nhy nhng kt qu chm v t tin. Phn ng khuch i gen PCR: K thut ny c u im l rt nhy v c hiu. C th chn on sm HIV giai on "ca s" hay tr s sinh nhng rt t tin. 8.5.3. Pht hin khng nguyn Khng nguyn P24 c trong huyt thanh, dch no tu v c rt sm sau khi nhim HIV. Thng dng k thut ELISA hay RIA pht hin khng nguyn ny. Tuy nhin k thut ny khng dng cho xt nghim sng lc v nhy v c hiu thp. 8.5.4. Cc xt nghim huyt hc v min dch Hng cu, bch cu, tiu cu gim. T bo lympho TCD4 gim di 400/mm3 (bnh thng 800 1200/mm3). T l lympho TCD4/ lympho TCD8 di 1 (bnh thng = 2). globulin mu tng.

Phng bnh: + Vaccin: Hin nay c nhiu loi vaccin ang giai on th nghim.
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+ Phng bnh ly nhim qua ng mu: m bo tt cng tc an ton trong truyn mu v cc sn phm ca mu. Khng tim chch ma tu. An ton trong tim chch v cc can thip y t. + Phng ly nhim HIV qua ng tnh dc. + Phng ly nhim HIV t m sang con: Khuyn nhng ph n nhim HIV khng nn c con, nu c thai th nn m . iu tr: + Thuc ngn cn s nhn ln ca HIV: Retrovirr, AZT, Interferon. Hin nay ng dng tr liu phi hp 3 thuc. + Chng nhim trng c hi. + Tng cng min dch bng g globulin v cc thuc kch thch min dch.
CU HI LNG GI
1. Trnh by c im sinh hc v kh nng gy bnh ca virus cm. 2. Nu phng php chn on VSH virus cm. 3. Nu nguyn tc phng v iu tr bnh cm. 4. K tn 2 khng nguyn quan trng nh typ virus cm. 5. Gii thch ngun gc cc dch cm. 6. Trnh by c im sinh hc v kh nng gy bnh ca virus quai b. 7. Trnh by c im sinh hc v kh nng gy bnh ca virus si. 8. Trnh by c im sinh hc ca virus vim no Nht Bn B. 9. Nu 2 th bnh do virus vim no Nht Bn B. 10. Nu c im dch t v phng php phng bnh virus vim no Nht Bn B. 11. Trnh by c im sinh hc ca virus Dengue. 12. Trnh by 2 th bnh Dengue xut huyt. 13. Nu nguyn tc phng v iu tr bnh st xut huyt Dengue. 14. Trnh by c im sinh hc v kh nng gy bnh ca virus bi lit. 15. Trnh by c im sinh hc v kh nng gy bnh ca Rotavirus. 16. Trnh by c im sinh hc v ng ly truyn ca cc virus vim gan A, B, C. 17. K tn cc khng nguyn v khng th quan trng ca virus vim gan B v ng dng trong chn on v theo di iu tr ca cc khng nguyn, khng th . 18. Trnh by c im sinh hc v kh nng gy bnh ca virus Varicella zoster. 19. Trnh by c im sinh hc v kh nng gy bnh ca virus di. 20. Trnh by c im sinh hc ca HIV. 21. Trnh by s xm nhp ca HIV vo t bo v s nhn ln ca virus. 22. Nu phng php chn on vi sinh v cc bin php phng nhim HIV.
TI LIU THAM KHO CHNH

1. V Triu An, Homberg, J.C. (1998). Min dch hc, Nh xut bn Y hc H Ni. 2. Bergeys Manuals of Microbiology (1994). 3. B mn Cng nghip Dc, Trng i hc Dc H Ni (2001). K thut sn xut Dc phm, Tp I, Trung tm Thng tin Th vin, i hc Dc H Ni. 4. B mn Ho sinh Vi sinh, Trng i hc Dc H Ni (1999). Vi sinh hc, Trung tm Thng tin Th vin, i hc Dc H Ni TV HD H Ni. 5. B mn Min dch Sinh l bnh, Trng i hc Y, H Ni (2003). Min dch hc, Nh
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xut bn Y hc, H Ni. 6. B mn Vi sinh vt, Trng i hc Y H Ni (2003). Vi sinh hc Y hc, Nh xut bn Y hc. 7. B Y t (2002). Dc in Vit Nam, Nh xut bn Y hc. 8. B Y t (2005). Hng dn s dng khng sinh, NXB Y hc, H Ni. 9. W. Crueger, A. Crueger. Biotechnolgia, Mezoegazdasgi Kiad, Budapest, 1987. 10. Nguyn Ln Dng, Nguyn nh Quyn, Phm Vn Ty (2002). Vi sinh vt hc, Nh xut bn Gio dc, H Ni. 11. H Hunh Thu Dng (2002). Sinh hc phn t, NXB Gio dc. 12. Elizabeth Forget (2003). Ti liu hi tho chuyn nm gy bnh, i hc tng hp Paris V. 13. Dr. Grg Jen (1973). Ipari Mikrobiologiai Gyakorlatok, Tanknyv Kiad, Budapest. 14. Phm Thnh H (2004). Di truyn hc, NXB Gio dc. 15. B. Hugo & A. D. Rusel. Pharmaceutical Microbiology, p. 428450, Blackwell Scientific Publications, Oxford, 3th Edition, 1983. 16. Yoichi Matsubara (2005). Healthcare in the Erea of Genomic Medicine, Paper on The 5th Science Council of Asia (SCA) Conference, May 1113, 2005, Daewoo Hotel, Hanoi, Vietnam. 17. Janeway, CA. and Tranvers, P. (1997). Immunobiology: The Immune system in Health and Disease, 3rd edn. Garland Publishing, New York. 18. T Minh Kong (2004). C s cng ngh sinh hc v sn xut Dc phm, NXB Y hc, H Ni. 19. J.P. Larpent, J.J. Sanglier (1989). Biotechnologie des Antibiotiques, Masson. 20. Lsztity Radomir (1973). Biokmia, Tanknyv Kiad, Budapest. 21. L nh Lng, Phan C Nhn (1997). C s di truyn hc, NXB Gio dc, H Ni. 22. Trn Xun Mai (2004). Vi nm y hc. 23. Dr. Sevella Bla (1988). Biomrnki M veletek II, Tanknyv Kiad, Budapest. 24. E.B. Shirling D. Gottlieb (1966). Methods for characterozation of Streptomyces species, Int. J. Bacteriol., Vol.16 (3), 313 340. 25. Pr.Dr. C. Simon, Pr.Dr. W. Stille, Dr. Muennich D.. Korszer Antibiotikus Kezels, Medicina Koenyvkiad, 1974, Budapest. 26. Szab I. Mihly (1980). ltalnos mikrobiolgia Vol. 14, Tankoenyvkiad, Budapest. 27. Tadeusz Korzebski, Zuzanna Kevsuk Gendifer, Wzodziniere Kuruzowicz (1967). Antibiotics, p. 11, 1167, PWN Polish Sicentific Publishers, Warsava. 28. Phm Vn Ty. Min dch hc (2001), Nh xut bn i hc quc gia H Ni. 29. Vin Y hc lm sng cc bnh nhit i (2001). Ti liu o to chuyn ngnh truyn nhim. 30. Moselio Schaechter, Ph. D.; N. Cary Engleberg, M.D.; Barry L. Eisenstein, M.D.; Gerald Medoff, M.D., (1999). Mechanisms of Microbial Disease, Lippincott Williams & Wilkins. 31. B.R. Champ, E.Highley, A.D.Hocking and J.I. Pitt (1991). Fungi and Mycotoxins in Stored Products, ACIAR Proceedings, No. 36.
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Chu trch nhim xut bn: Ch tch HQT kim Tng Gim c NG TRN I Ph Tng Gim c kim Tng bin tp NGUYN QU THAO Chu trch nhim ni dung: Ch tch HQT kim Gim c Cng ty CP Sch HDN TRN NHT TN
Bin tp ni dung v sa bn in: NGUYN HNG NH Trnh by ba: INH XUN DNG Ch bn: TRNH THC KIM DUNG
In 1300 bn, (Q:..79...), kh 19 x 27 cm, ti Cng Ty Phn In Phc Yn a ch: ng Trn Ph, th x Phc Yn, Vnh Phc. S KKH xut bn: 922-2008/CXB/6-1873/GD. In xong v np lu chiu thng 12 nm 2008.
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Vi Sinh Vat Hoc (Duoc Si)

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Vi Sinh Vat Hoc (Duoc Si)

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LI GII THIU

FAD FADH2 FHA FMN FTA

Flavin adenin dinucleotid dng oxy ho Flavin adenin dinucleotid dng kh ho

1. VI NT V LCH S PHT TRIN CA VI SINH VT HC

2.1. Kch thc ca vi sinh vt rt nh b

2.2. Vi sinh vt hp th nhiu, chuyn ho nhanh, vng i ngn

2.3. Vi sinh vt sinh trng nhanh, pht trin mnh

2.4. Vi sinh vt thch ng mnh v d pht sinh bin d

2.5. Vi sinh vt phn b rng, chng loi nhiu

3. V TR CA VI SINH VT TRONG SINH GII

4. PHN B VI SINH VT TRONG T NHIN

4.1. Vi sinh vt trong t v trong nc

4.2. Vi sinh vt trong khng kh

4.3. Vi sinh vt c th ngi lnh

5. CC SN PHM DC DNG C NGUN GC T VI SINH VT

HNH THI V CU TO T BO CC VI SINH VT PROCARYOTA

1.1. Hnh thi v kch thc vi khun

Hnh 1.2. S cu to t bo vi khun

Hnh 1.7. Cu trc ca mng t bo cht

Hnh 1.13. Hnh dng cc chui bo t ca x khun

2.1. Cc c im hnh thi ca x khun

2.3. Mt s kho phn loi v cc c im phn loi ca x khun

3. NHM CC VI KHUN NGUYN THU

Hnh 1.16. Rickettsia rickettsii trong t bo vt ch

DINH DNG VI SINH VT

1. CC CHT DINH DNG CA VI SINH VT

1.1. Ngun dinh dng carbon ca vi sinh vt

1.2. Ngun dinh dng nit ca VSV

1.3. Ngun dinh dng khong ca VSV

1.4. Cc cht sinh trng

2. THNH PHN T BO V DINH DNG CA VI SINH VT

2.1. Thnh phn nc v mui khong

3. C CH VN CHUYN CC CHT DINH DNG VO T BO VI SINH VT

Hnh 2.1. Vn chuyn ch ng cht dinh dng vo t bo

Hnh 2.2. Vn chuyn kt hp phosphoryl ho

TRAO I CHT V TRAO I NNG LNG

1. CC KHI NIM CHUNG

H O ribose adenosin AMP ADP AT P

phosphat phosphat phosphat

2. CC CON NG OXY HO SINH HC PHN GII HYDRATCARBON CC VI SINH VT D DNG

2.1. Phn gii glucose

Hnh 3.1. Con ng ng phn Embden-Mayerhof-Parnas

L v tr cung cp lin kt phosphat cao nng (~P). Phn ng tng qut:

O H G-6-PDH H O OH OH OH H H 6-Phosphoglucono-1,5-lacton Glucose-6-phosphat H O H H2O, Mg

Lactonase COOH C O H C H hoc H H O

CH2O P OH Dehydratase H HO H O 2+ Fe , GSH H OH H O H2O OH H 6-Phosphogluconat

O COOH C O CH3 Pyruvat CH C O CH2 O P Pyruvat

Hnh 3.3. Con ng EntnerDoudoroff (ED)

2.2. Phn gii cc loi ng khc

Galactose sau khi bin thnh Glucose6P cng i vo EMP:

Phosphoglucose uridin transferase

Nh vy, trong trng hp ny khng cn s dng nng lng t ATP m vn c th to ra c glucose1phosphat.

3.1. Qu trnh oxy ho pyruvat

+ 1. Pyruvat + CoA + NAD

AcetylCoA + NADH2 + CO2

COO + + (+) + H + H + () C C O () COO CH3 C OH CO2

H CH3 C C carboxylase OH acetaldehyd hot ng

CH2 CH2 S SH CH2 CH2 HOOC O (CH2)4 CH S C CH3

CH S acid lipoic O C (CH2)4 Enzym

Hnh 3.4. Oxy ho pyruvat nh pyruvat dehydrogenase

3.2. Chu trnh Krebs

NADH 2 O CH C 3 CO2 Acetyl-CoA S CoA

H C COOH 2 Citratsynthase H C COOH Citrat O H C COOH 2 Aconitase HO 2 H C COOH 2 C COOH H C COOH HO 2