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LI M U

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Vi sinh vt c khp ni xung quanh chng ta nh trong nc, khng kh, thc phm v hin din c trn c th chng ta. c bit l thc n v nc ung l con ng chnh vi sinh vt xm nhp v gy bnh cho con ngi.. Vi tnh hnh an ton v sinh ang din ra ht sc phc tp. Song song , vn v sc khe cng l mt vn mang tnh thi s nng hi. Chnh v th m khng t ngi t cu hi: Liu trong thi i th trng cnh tranh khc lit nh hin nay th cc doanh nghip c thc s ch trng n sc khe ngi tiu dng khng hay li nhun s c t ln hng u? Hin nay, ng c thc phm tr thnh mi quan tm ca ton x hi. Cc v ng c thc phm th c rt nhiu nguyn nhn nh do: ha cht, bn cht thc phm cha sn mt s cht c, Nhng quan trng hn ht vn l t cc vi sinh vt, trong cc vi khun l mt trong nhng nguyn nhn chnh gy ng c. Nhm mc ch tm hiu ngun gc pht sinh v nhng tc hi m cc vi khun ny gy ra cng nh cc bin php phng nga v cha tr... c bit l tm hiu cc phng php phn tch nhn bit v pht hin chng, y ng thi c th xem l mt bin php hu hiu kim tra an ton ca thc phm.

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Chng 1. TM HIU CHUNG V NG C THC PHM 1.1.Tnh hnh ng c thc phm trong nc v trn th gii 1.1.1. Tnh hnh ng c thc phm trn th gii T nm 1973 n nm 1987, theo thng k ca C quan Kim sot bnh tt (Centers for Disease Contrl : CDC) ti Atlanta (Hoa K), c tnh c khong 500 trn dch bnh do thc n gy nn, nh hng cho khong trung bnh 16.000 ngi mi nm. Tuy nhin nhng con s ny cha c nh gi mt cch y , cha phn nh tht s tm quan trng ca vn . T l mc bnh hng nm l bao nhiu th khng r, nhng theo thng k ca cc c quan y t Hoa k, ch ring ti Hoa K, c tnh c khong chng 6.500.000 n 81.000.000 lt bnh mi nm. chu, nhiu trn dch ng c thc n gn y c ghi nhn ti Nht v iu ny lm cho nhiu quc gia khc phi lo ngi[1]. T thng 3 n thng 5 nm 2010, mt bin c ng c do vi khun E coli nhim t ci Romaine lettuce xy ra ti Michigan, New York, Tennessee v Pennsylvania Hoa K. Th phm l mt chng E coli mi, l E.coli 0145. y l mt chng loi him thy nn t c cc nh khoa hc quan tm n. E coli 0145 cng gy bnh tng t nh E coli 0157:H7. Bin c trn lm cho vi chc ngi ng bnh. Vo trung tun thng 5, 2011, Chu u chn ng v dch bnh ng c thc phm do E.coli 0104, mt chng lm hy hoi xut huyt nim mc rut Enterohemorrhagic E.coli EHEC. y l mt loi rt him thy t xa nay. Ngi ta nghi rng ngun ly nhim l t da leo hu c organic sn xut t vng Almeria v Malaga, thuc Ty Ban Nha v c nhp vo cc quc gia Chu u. Tnh n 30/5/2011, c l quc gia b nng nht vi gn 400 ngui bnh v c 14 t vong v an Mch c 11 t vong. Ngoi ra, dch bnh cng c thy xut hin ti Thy in, Anh Quc, H Lan[2] Vi khun l mt trong nhng nguyn nhn gy ng c thc phm nhiu nht. Ti nc Anh, trong 2000 ca b ng c thc phm ring l do vi khun th Campylobacter jejuni chim 77,3%, Salmonella 20,9%, Escherichia coli SVTH: Phm TH Hu Hu_11SHLT
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Escherichia coli O157:H7 1,4%, cc vi khun cn li gy ra t hn 0,1% s ca[16]. Triu chng ng c thc phm do vi khun thng ch xut hin sau 1272 gi hoc hn na sau khi n thc n b nhim khun[3]. Hoa K: theo cc s liu ca FoodNet t nm 1996 n 1998, CDCP c tnh rng hng nm c 76 triu ca ng c thc phm (26 nghn ca trn 10 vn dn), trong : 325 nghn nhp vin (111/10 vn dn); 5 nghn cht (1,7/10 vn dn). Tn tht gy ra bi s st gim nng sut cng chi ph iu tr nm 1997 l 35 t M kim[3]. Php: hng nm c 75 vn ca ng c thc phm (1.210 ca/10 vn dn), trong : 7 vn ca cp cu trong tnh trng nguy kch (113 ca/10 vn dn); 113 nghn ca phi nhp vin (24 ca/10 vn dn); 400 ngi cht (0.9 ngi/ 10 vn dn)[3]. c: hng nm c tnh c khong 5,4 triu ca ng c thc phm, c th: 18 nghn ca nhp vin, 120 ca t vong, 2,1 triu ca phi ngh lm v ng c, 1,2 triu cuc hi chn gia cc bc s, 30 vn toa khng sinh[3]. 1.1.2. Tnh hnh ng c thc phm trong nc Ti thnh ph H Ch Minh, vo thng 5/1996, mt v ng c thc n sau ba tic lin quan c ghi nhn ti trng tiu hc Bu Sen, qun 5, 818 hc sinh tham d, 164 em b ng c. Qua xt nghim vi sinh tm thy Staphylococcus aureus trong tht g, bnh m v c trong cht nn ca bnh nhn[1]. Vit Nam nhiu v ng c thc phm hay xy ra, c bit l ng c tp th, ri nhiu vo i tng cng nhn (khi n, ung ti cc bp n tp th khng m bo v sinh, an ton cht lng thc phm). Theo mt thng k nm 2008, mi nm Vit Nam c khong 250- 500 ca ng c thc phm vi 7.000 - 10.000 nn nhn v 100 - 200 ca t vong[3]. C 8 triu ngi ng c thc phm mi nm ti Vit Nam. y l cng b ca T chc Y t th gii (WHO) v tnh hnh ng c thc phm ti Vit SVTH: Phm TH Hu Hu_11SHLT
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Nam. Tuy nhin ch c 8.000 ngi c thng k, pht hin, bng 1% s ngi ng c thc phm trn thc t[4].

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Chng 2: TM HIU CC VI KHUN GY NG C THC PHM 2.1. T cu khun Staphlococus aureus Staphylococcus aureus l vi khun hnh cu hoc hnh thun. Trong bnh phm vi khun hp tng i hoc m nh. Vi khun bt mu Gram dng. Vi khun khng di ng, khng sinh nha bo, thng khng c v. Thng gp do n thc n giu m b nhim vi khun t cu nh: tht, c, trng, sa, cc loi sp... Vi khun t cu c nhiu trn da, hng khi b vim nhim, v c trong khng kh, nc... nn qu trnh ch bin v bo qun khng hp v sinh rt d nhim cc vi khun ny vo thc phm. n thc n c nhim t cu hoc c t ca chng u c th b ng c. Bnh thng, triu chng xut hin sm trong 30 pht n 4 gi sau khi n. Ngi bnh thng nn thc n va n xong, i ngoi nhiu ln phn ton nc, mt mi, c th c au u hn m nu nhim phi c t ca t cu. Bnh khng c iu tr kp thi d t vong do mt nc v in gii. iu tr tch cc, bnh thng khi nhanh v phc hi tt[5]. 2.2. Vi khun E.coli E.coli - Vi khun ny hin din mt cch t nhin trong rut ca chng ta cng nh ca th vt. Vi khun E.coli thng nhim vo t, nc, tht, rau, qu, sa t phn ca ng vt. C c hng trm chng huyt thanh (serotypes) E.coli. a s u l nhng chng hin, tuy nhin cng c vi chng rt d, chng hn nh E.coli 0157:H7 c th c tm thy trong rut v trong phn ca cc loi gia sc, c bit l trong phn b. Vi khun ny c xc nh ln u tin vo nm 1982 ti Hoa K nhn bin c ng c thc phm do hamburger gy ra. T vi khun E.coli 0157:H7 cn c tn l vi khun ca bnh hamburger. c t verocytoxin ca E.coli 0157:H7 lm dung huyt (hemolysis), hy hoi nim mc rut gy tiu chy c mu, lm h thn v ng thi lm gim lng nc tiu. Khoa hc gi y l hi chng HUS (Hemolytic Uremic Syndrome), rt nguy him v c th cht, bng khng th cng cn phi c lc thn (renal dialysis) sut i[2]. SVTH: Phm TH Hu Hu_11SHLT
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Khi nhim vi khun ny thng qua ng thc n, th ngi b s bnh t 2-20 gi. Ngi ng c thy au bng d di, i phn lng nhiu ln trong ngy, t khi nn ma. Thn nhit c th hi st. Trng hp nng bnh nhn c th st cao, ngi mi mt, chn tay co qup m hi. Thi gian khi bnh vi ngy[3]. - E .coli: c 3 nhm E. coli gy a chy xm nhp vi hi chng l: + EIEC (E. coli Entero-invasive) bnh xy ra tr em, ngi ln, vi hi chng l nh l trc trng. + EPEC (E. coli Entero pathogenic), trc y, n l tc nhn gy cc dch a chy nht l tr em trong cc nh tr, mu gio nhng hin nay n t phn lp c bnh nhn a chy. iu tr khng sinh l cn thit tr nh. + EHEC (E. coli Entero Hemorragique colitis): gy a chy phn nhy mu[11]. 2.3. Vi khun Listeria Listeria monocytogenes l mt tc nhn gy bnh listeriosis rt nguy him, gy t vong cao ngi nh nhim trng mu, sy thai ph n, vim mng no, gy t vong thai nhi. T l t vong khi mc phi bnh listeriosis khong 20%. S nhim c L. monocytogenes gy ra bi mt s tc nhn gy c trung gian, trong c listeriolysin O (LLO) l c t quan trng nht c tit ra bi mm bnh[14]. Listeria monocytogenes gp trong rut ca ng vt v trong t ct. Vi khun Listeria c th nhim vo trong cc loi rau ci ti. c bit hn na l n c th m thm tng trng trong t lnh nhit 40C. Listeria cng c th nhim vo tht ngui, jm-bng, pho mt, sa ti nu khng c hp kh trng trc khi bn[2,6]. 2.4. Vi khun Salmonella Ging Salmonella l vi sinh vt ng rut Enterobacteriaceae c hn 2400 kiu huyt thanh. Salmonella l trc trng Gram m, kch thc khong SVTH: Phm TH Hu Hu_11SHLT
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0,6 - 2,0 mm, hiu kh, c th di ng, khng to bo t, sinh hi ln men dextrosa, sinh kh ihyrosunfua[7]. Salmonella kem khang vi iu kin bn ngoai, bi pha huy bi qua trinh tit trung bng phng phap Pasteur, tia bc xa va un si nu ki. Tuy nhin Salmonella co th sng sot trong mt thi gian dai cac thc phm kh va p lanh. Do o khi lam tan thc phm ng lanh vi khun nay d phat trin tr lai. Chung phat trin tt nhit 60C 420C, thich hp nht 350C 370C, pH t 6 9 va thich hp nht pH = 7,2 . nhit t 180C 400C vi khun co th sng n 15 ngay. un si nc 600C trong 10 phut hay 1000C trong 2 phut a dit c hu ht Salmonella. Chung b tiu dit bi phenol 5%, cloramin1% v clorua thy ngn 0,2% trong 5 pht. Nui cy d dng v tng trng nhanh trong mi trng n gin, nhng mi trng ny phi cht dinh dng, vi hot nc Aw = 0,93. Vi khun ny khng gy mi v kh chu cho thc phm. Sng tt mi trng bn ngoai, co th sng c ca vi iu kin bao quan, p mui, it nc. No co kha nng chiu c khang sinh. Trong iu kin nhit . S xm nhim Salmonella vo c th vt ch v gy bnh c thc hin ch yu qua ng tiu ho vi biu hin ph bin nht l gy tiu chy, i khi l thng hn v ph thng hn. Salmonella chu yu gy bnh bng ni c t. Ni c t chu c nhit si v khng b phn hy bi protease, tnh khng nguyn yu v khng sn xut c thnh khng nguyn. Tri li vi ngoi c t. gy bnh, Salmonella xm nhp vao c th theo ng tiu hoa do thc n, nc ung bi nhim bn, s lng gy bnh chi khoang 10 5 n 107. Cc chng Salmonella thng sn sinh ra 1 entertoxin c bn cht lipopolysaccharide vn c kh nng tc ng n nhiu m khc nhau, n cc chc nng ca m. Tuy nhin trong trng hp nhim c thc phm SVTH: Phm TH Hu Hu_11SHLT
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cht c ny ch c tc dng khi n c gii phng vo trong rut t nhng vi khun sng v ang trong pha sinh sn. Khi n cc bo t sng th c th sinh bnh song khi n cc vi khun b cht do nhit th khng b nh hng g. Sau khi i vao ng tiu hoa, vi khun bam vao nim mac rut non ri xm nhp qua nim mac vao cac hach mac trn rut. y, chung nhn ln ri qua h thng bach huyt va ng ngc i vao mau, luc nay du hiu lm san bt u xut hin. T mau, vi khun n l lach va cac c quan khac: + Ti mang Payer, vi khun tip tuc nhn ln. + Ti gan theo mt xung rut ri c ao thai qua phn. + Ti thn, mt s vi khun c ao thai ra ngoai theo nc tiu. Salmonella gy bnh bng s xm nhp cua ban thn vi khun pha huy t chc t bao bng ni c t cua Salmonella khi bi cht. Ng c do Salmonella cn co 2 iu kin: + Thc n phai nhim mt lng ln vi khun sng, vi tinh cht gy ng c cua vi khun rt yu + Vi khun vao c th phai tit ra mt lng ln c t[12]. Salmonella c th c tm thy trong phn ca cc loi vt v gia cm. Ra v rn v cc loi b st cng thng c mang vi khun Salmonella. Tht b, tht heo nht l tht g, trng g, sa ti v cc loi thy sn nh c, tm, s c, rau ci u c th b nhim khun salmonella[2]. y l tnh trng hay gp trong cc v ng c ti bp n tp th nh cc trng hc bn tr, cc x nghip sn xut, cc bui lin hoan hay l ci... Vi khun gy ng c a s l nhm vi khun ng rut, kh nng gy bnh ca nhm ny yu nn gy bnh thng phi c mt lng ln thc n. Ng c thc phm loi ny thng xy ra trong vng vi gi n mt ngy sau khi n cc thc phm b nhim ny.

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Triu chng: Khong 12-14 gi sau khi n thc phm c cha vi khun, nn nhn s c cc triu chng: au bng, nn, tiu chy, ton thn b lnh ri st, suy nhc c th. Nu khng cp cu kp thi c th dn n t vong[3]. 2.5. Vi khun Shigella Shigella gy bnh l trc khun ngi, y l mt bnh truyn nhim c th gy thnh cc v dch a phng. Thng tn c hiu khu tr rut gi, trn lm sng biu hin bng hi chng l vi cc triu chng: au qun bng, i ngoi nhiu ln, phn c nhiu m nhy v thng c mu. Shigella gy bnh bng c ch xm nhp vo t bo biu m ca nim mc rut v nhn ln vi s lng ln trong t chc rut. Cc Shigella u c ni c t. Ring trc khun Shiga cn c thm ngai c t c bn cht l protein. Ni c t Shigella nh khng nguyn thn, c c tnh mnh nhng tnh khng nguyn yu. Tc dng chnh ca ni c t l gy phn ng t rut. Ngoi c t ca trc khun Shiga khng ging nh c t rut ca Vibrio cholerae 01 v ETEC, hot tnh sinh hc ch yu ca ngoi c t trc khun Shiga l tc dng c i vi t bo. Vit Nam, Shigella gy bnh l trc khun thng gp nht l nhm B (Shigella flexneri) v nhm A (Shigella dysenterae). Bnh ly theo ng tiu ha, do n ung phi cc thc n, nc ung b nhim khun. Rui l vt ch trung gian truyn bnh. Ngi lnh mang vi khun v ngi bnh ng vai tr quan trng gy dch. Dch thng xy ra vo ma h. Nhim khun Shigella, hay Shigellosis, l bnh nhim khun ng rut gy ra bi mt trong nhiu loi vi khun Shigella. Cc loi vi khun ny c th ly truyn qua thc n v nc ung cng nh thc phm km v sinh. Vi khun xm nhp mng trong rut, c th dn n triu chng tiu chy, st hay chut rt[8].

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Shigella - Ly truyn t nhng ngi bin ch thc n khng chu ra tay cho k lng trc khi s m vo rau ci v thc phm ti sng. Khun Shigella c th c tm thy trong tht g, trong cc da salade v trong sa. Triu chng pht hin ra sau khi n mt vi ngy l au bng qun tht, st nng v tiu chy thng c mu. Khi bnh sau 5-7 ngy. Trng hp nng c th thy cc tr em di 2 tui. Cc chu c th b ng kinh v co git. Mt s ngi b nhim m khng b bnh g ht nhng h li c th ly nhim cho cc ngi khc[2]. 2.6. Vi khun gy bnh thuc ging Vibrio Vibrio l nhng vi khun hnh que hi cong, di ng nh mt lng mt u, hiu kh hoc k kh ty tin. Hu ht c phn ng oxidase v catalase dng tnh, ln men ng nhng khng sinh hi, chuyn natri thnh nitrit, cn Na+ (mui NaCl) cho s pht trin. Chng thng xut hin cc vng nc ven bin, khng thch hp vi mi trng thiu mui[9,11]. Vibrio cholerae l chng vi khun gy ra nhng trn dch t trn ton th gii. Dch t gy ra do V. cholerae thng lan truyn nhanh qua ng nc, gy nhim thc phm v truyn nhim qua con ngi khi iu kin v sinh km. Cc loi rau sng c ti hoc ra bng nc nhim bn s gy nhim khun vo thc n[14]. Vi khun Vibrio cholerae c thy trong bin, c, tm tp, nghu s, nc bn... Cc trng hp l lt thng to iu kin thun li d lm ly lan dch bnh cholera. Vibrio cholerae gy bnh t, khi b nhim au bng qun tht, i ma, tiu chy d di c chc ln trong mt m, phn lng v trng c nh nc vo go. C th b suy xp rt nhanh v mt nc ko theo mt cht in gii v bi thn. Khng cha tr kp thi s cht[2,6]. 2.7. Vi khun Clostridium perfringens Vi khun Clostridium perfringens (C. perfrin-gens) l mt loi vi khun thng thy trong t, nc thi, rut con ngi v ng vt. SVTH: Phm TH Hu Hu_11SHLT
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Vi khun C. botulinum ch pht trin trong iu kin hon ton khng c khng kh m thi. Ngi ta gi chng l vi khun ca nh n (The Cafeteria germ) v chng thng hin din trong cc thc phm ngui lnh ca cc ca hng n ung. Vic nu nng i lc cng khng th dit ht mm bnh c, mt s vi khun c th vn cn sng st, n ly lan t ngi ch bin thc phm sang thc phm, tip tc sinh si ny n pht trin v sn xut ra c t. n phi nhng thc n va k, c t ca vi khun C. botulinum s gy ra bnh Botulism rt nguy him nh nut kh, n ni kh khn, x m mt, t lit dn dn h h hp v c th nhn thy c 2 nh cng mt lc (double vision). C. perfringens thng c tm thy trong cn tin trng hc, bi n pht trin mnh cc loi thc phm c phc v vi s lng ln, v bo qun nhit phng. Cc triu chng nhim c xut hin t 8-24 gi sau khi n, c th ko di t mt n vi ngy[2,6,8]. 2.8. Cc vi khun gy ng c khc Calicivirus - Virut ny cng thng gy ng c thc phm, nhng t khi c ngi ta nh bnh mt cch chnh xc c. Triu chng l au bng v i ma d di nhiu hn l tiu chy. Bnh thng dt sau 2-3 ngy. Virut c tm thy trong cht nn ma v trong phn ca ngi bnh. Khc vi cc mm bnh thng gp u c ngun gc t th vt, Calicivirus thng ly truyn t ngi ny sang ngi khc qua vic s m, chun b v ch bin mn n[6]. Cryptospora v Giardia lamblia - y l hai loi k sinh trng thuc nhm nguyn sinh vt c trong phn sc vt v c th nhim vo ngun nc (k c nc ging) v rau ci... Vibrio vulnificus - Gp nhng vng ven bin. Ngi c th b nhim qua cc vt try trn da t nc bin, hoc do n phi nhng loi bin, nh nghu s, c cha vi khun V. vulnificus... au bng, nn v tiu chy l nhng triu chng chnh. nhng ngi gi c hoc nhng ngi c sc min dch km, h c th b nhim trng huyt, ni mn nc ngoi da, gim huyt p ng mch v cht v b sc[6]. SVTH: Phm TH Hu Hu_11SHLT
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Campylobacter jejuni - thng hin din trong rut ca cc loi gia sc v gia cm... Phn c th nhim vo ngun nc v cc loi thc n, nh tht g, sa v rau ci. C th ni ng c thc phm do vi khun Campylobacter l bnh tiu chy thng hay xy ra nht. Triu chng ca bnh pht hin ra sau khi n t 2 n 5 ngy, v thng l au bng, nn v tiu chy c th c mu. Bnh s dt sau 1 tun l[6].

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Chng 3: P DNG PHN TCH PHN TCH BNG NUI CY MT S VI KHUN GY NG C THC PHM 3.1. Xc nh vi khun gy ng c mu th l thc phm Mu xc nh vi ly t cc loi thc phm nghi ng gy ng c, c thu v x l nghin nh, trch ly, pha long nui cy phn lp vi khun gy ng c trong thc phm. - E.coli: tht, sa, rau qu - Salmonella: tht (nht l tht g), trng g, sa ti v cc loi thy sn - shigella: tht g, sa - Staphylococcus: tht, c, trng, sa, sp - Vibrio: thy sn: c, tm, cua 3.1.1. Xc nh Coliforms[15] Phng php: tham chiu theo phng php NMKL 44 n bn ln 4, 1995 Nguyn tc Da vo s ln ln men ng lactose mi trng thch hp (thch Violet red bile) 370C trong 24 gi. Sau c khng nh li trong mi trng canh Brilliant Green Bile Salt. Coliforms s sinh kh trong mi trng ny 370C trong 24 gi. Mi trng v thit b - Dung dch Salin Pepton - Thch Violet red bile (VRBL) - Canh Brilliant Green Bile Salt (BGBL) - Thch Tryptone Soya (TSA) - T n 3710C Quy trnh a: chuyn 1ml dung dch dung dch mu sau khi pha long cho vo a petri v trng, s dng hai nng pha long lin tip. vo mic a khon 5ml mi trng TSA 450C. Sau cho mi trng ng hon ton thn 10-15ml mi trng thch VRBL 450C SVTH: Phm TH Hu Hu_11SHLT
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Nui : Cc a c lt ngc v trong 243 gi 3710C c kt qu m cc a c s khun lc di 100 sau 24 gi nui cy. Khun lc coliforms c mu ta, ng knh 0.5mm, i khi c bao quanh bi mt vng hi do ta. Tnh gi tr trung bnh t cc pha long qui v s coliform trong 1g mu. Khng nh. Cy ring t nht 5 khun lc nghi ng ca mi loi vo cc ng nghim cha mi trng canh BGBL c ng Durham , 370C trong 24 gi. Phn ng c coi l dng tnh khi c s to kh d ch mt trong 5 ng nghim trn. 3.1.2. Xc nh Escherichia coli (E.coli)[15] Phng php: tham chiu theo TCVN 5287, n bn ln 2, 1994 Nguyn tc Cy mt lng dch mu vo mi trng tng sinh (canh Brilliant green bile lactose), chn cc khun lc in hnh trn mi trng chn lc (EMB) cc php th sinh ha ph hp (nghim php IMViC) Mi trng v thit b - Dung dch Saline (SPW). - Canh Brilliant green bile lactose BGBL. - Thch Eosin Methylene Blue Lactose (EMB) - Canh Methyl Red Voges Proskauer (MR-VP) - Canh Tryptone (hoc peptone) - Thch Simmons Citrat - Thuc th Methyl red 0,2%, -naphtol 5%, kovacs. - Dung dch KOH 40% - T m 3710C, 4410C Quy trnh Tng sinh: Cy 1ml dch mu nng 10-1 vo ng nghim cha 5ml mi trng tng sinh (BGBL), 44,00,50C 24 gi.

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Cy phn lp: Sau khi tng sinh cy dch mu t ng nghim c phn ng dng tnh (mi trng chuyn c v sinh hi) sang mi trng EMB, 371/24 gi. Trn mi trng EMB: khun lc mu tm, nh kim, trn, b iu, ng knh khong 0,5mm.

Hnh 3.1. S xc nh E.coli 0157 trong thc phm Khng nh Chn ch nht 2 khun lc in hnh trn mi trng chn lc sang mi trng thch khng chn lc (TSA) 3710C trong 19-24 gi. Kt qu th nghim sinh ha E.coli ph hp: - Indol - Methyl red : (+) : (+)

- Voges proskauer : (-) SVTH: Phm TH Hu Hu_11SHLT

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- S dng citrat : (-)

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3.1.3. Xc nh Salmonella[15] Phng php: tham chiu theo phng php NMKL 71 n bn ln 5. Nm 1999 Nguyn tc Phng php ny ch dng nh tnh pht hin hay khng pht hin Quy trnh kim tra Salmonella bt buc phi qua bn gia on: tin tng sinh, tng sinh, phn lp v khng nh. Mi trng v thit b - Canh Peptone m (BPW). - Canh Rappaport-Vasiliadis soy peptone. - Thnh Brilliant Green Phenol Red Lactose Sucrose (BPLS) - TSI, TSA - Canh Lysine decarboxylase, Urea phenol red, Manitol Phenol red base - B iu nhit 0.20C - T m 3710C Quy trnh Ly mu: Ly mu vng b mt cng rng cng tt Tin tng sinh: trn 25g mu vi 225ml nc m peptone m, ng nht nht bng my dp mu. 370,10C t 18 n 24 gi Phn lp: t mi trng sinh cy chuyn khun dch ln b mt mi trng phn lp XLD sao cho c th to c nhng khun lc tch ri. Lt ngc a 370,20C trong 243 gi. Trn mi trng XLD khun lc Salmonella in hnh trong, hi nhum do s thay i ca cht ch th trong mi trng, phn ln c tm en. Bao gi cng nhn thy mt vung mi trng ln hay nh. Khng nh Khun lc nghi ng c kim tra khng nh bng th nghim sinh ha: - Lactose - Sucrose : : (-) (-) - Mannitol - VP : : (+) (-)
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- Glucose - Urease - Indol : : : (+) (-) (-)

GVHD: Trn Th X
- LDC - ODC : : (+) (+) (-)

- Amygdaline :

3.1.4. Xc nh Clostridium perfringens[15] Phng php: tham chiu theo phng php NMKL 56 n bn ln 3. Nm 1994 Nguyn tc nh lng Clostridium bng cnh cy mt lng mu bit vo mi trng thch hp c cha ion Fe3+ v ion (S2O3)2- 370C trong 1-2 ngy. Mi trng v thit b - Thch iron sulphite - Dung dich mui peptone pha long Quy trnh Cy mu: Chuyn 1ml mu nng thch hp vo ng nghim. Sau , 12ml mi trng thch iron sulphite vo ng trn iu mu trc khi mi trng ng li. Sau khi mi trng ng thm 2-3ml mi trng thch iron sulphite ln b mt 3710C trong 28-48 gi. c kt qu m tt c khun lc en, xung quanh c qung en nhn vi nng pha long. 3.1.5. Xc nh Virio Cholerae v Virio parahaemolyticus[15] Phng php: tham chiu theo s tay phn tch vi sinh FDA , n bn ln 8, nm 1995 Nguyn tc Virio Cholerae v Virio parahaemolyticus c nui trong mi trng lng chn lc. t y dch khun c cy chuyn sang mi trng rn chn lc. Nhng khun lc ging Virio Cholerae v Virio parahaemolyticus c th nghim bng cc phn ng sinh ha. Mi trng v thit b SVTH: Phm TH Hu Hu_11SHLT
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GVHD: Trn Th X

- Thch Thiosulphate citrate Bile salt sucrose (TCBS agar) - Peptone kim cha 1% Nacl (APW) - TSA + 1,5 %Nacl - T m 3710C Quy trnh Tng sinh: Cn v trng 25 g mu thy sn vo mt ti PE v trng. Ct cc mu ln thnh cc mnh nh ri thm 225 ml nc pepton kim APW. Dp mu 2 pht, 3710C trong 6 gi v 16-24 gi Phn lp v c kt qu: t mi trng tng sinh cy ria vo mi trng TCBS thch 370C 1,00C trong 18 - 24 gi. Trn mi trng TCBS thch khun lc V. cholerae trn, ln c ng knh 2-3mm hi dt, mu vng (do vi khun ln men sacaroza), gia m v c vin m xung quanh. V. parahaemolyticus trn, ln c ng knh 3-4mm, mu xanh dng. Th nghim sinh ha Khun lc nghi ng trn TCBS thch cy chuyn sang mi trng khng chn lc TSA 1,5% NaCl. nhit 37,0 1,0oC trong 18 - 24 gi. Bng 3.1. Th nghim sinh ha s b Php th TSI Tinh duy ng Oxydase KOH Phn ng tiu biu ca Virio parahaemolyticus K/A -+ + + Phn ng tiu biu ca Virio Cholerae K/A -+ + +

Bng 3.2. Th nghim sinh ha khng nh Php th Tnh chu mn 0% NaCl 3% NaCl + +
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Phn ng tiu biu ca Virio parahaemolyticus

Phn ng tiu biu ca Virio Cholerae

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6% NaCl 8% NaCl 10% NaCl Ln men sinh axt t Sucrose Lactose Maninitol ONPG ADH LDC Urease + + + + + + + + +

GVHD: Trn Th X

3.1.6. Xc nh Staphylococci aureus c phn ng Coagulase dng tnh[15] Phng php: tham chiu theo phng php NMKL 66 n bn ln 3, 1999. Nguyn tc nh lng St. aureus Coagulase positive c thc hin bng cnh cy trang mt lng mu bit ln mi trng thnh Bard Parker, vi khun cho nhng khun lc t trng. V c xc nh bng phn ng coagulase. Mi trng v thit b - Dung dch Saline Peptone - Thch Trypton Soya - Thch Barid Parker - Canh Brain Heart Infusion (BHI) - Huyt thanh th - T n 3710C Quy trnh Cy trang 1ml mu vo 3 a a petri. Lt ngc cc a v 3710C trong 243 gi v 484 gi. c kt qu

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Sau 243 gi, trn mi trng Baird Paker khun lc Staphylococci aureus Coagulase positive c ng knh 1-1,5mm, mu en sng v li. Mi khun lc c qung sng rng 1-2mm bao quanh. Nhng khun lc in hnh s c nh du trn mt sau ca a v tip tc thm 24 gi na. Sau 48 gi Staphylococci aureus Coagulase positive 1,5-2mm c mu en, sng v li, quanh khun lc c mt vng c m hp, tip l mt vng sng trong rng 2-4mm . Mt vi chng St. aureus khng to qung sng trong bao quanh, vng c st khun lc cng c th khng c (khun lc khng in hnh). C hai loi khun lc ny iu c nh du mt sau ca a. Khng nh Cy 5 khun lc in hnh v khng in hnh sang mi trng TSA 3710C trong 24 gi. T mi trng TSA cy chuyn vi khun sang cc ng nghim c cha 0,3ml huyt tng th 3710C. kim tra s hnh thnh khi ng sau 1,3,6 v 24 gi. Kt qu - Dng tnh c s hnh thnh khi ng. - m tnh khng c s hnh thnh khi ng. 3.2. Xc nh vi khun gy ng c bng bnh phm hin i 3.2.1. Xc nh E.coli[11] Trong ng tiu ha E.coli chim t l cao nht trong s vi khun hiu kh (khong 80%). Tuy nhin, E.Coli cng l mt vi khun gy bnh quan trng, n ng u trong cc vi khun gy a chy, vim ng tit niu, vim ng mt; ng hng u trong cc cn nguyn gy nhim khun huyt. E.coli c th gy nhiu bnh khc nh vim phi, vim mng no, nhim khun vt thng. Nhng typ huyt thanh c kh nng gy bnh thng gp trn lm sng l: O111B4, O867B7, O126B16, O55B5, O119B4, O127B8, O25B15, O128B12. C ch gy bnh ca E. coli khc nhau ty loi: ETEC: gy bnh do ngoi c t LT, l loi c t rut ca V. cholerae. c t ny bm vo th th rut, lm gim hp thu Na+, tng tit nc v Cl-. SVTH: Phm TH Hu Hu_11SHLT
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EIEC: gy bnh do kh nng xm nhp vo nim mc i trng, c ch gy bnh ging vi khun l. EHEC: c ch cng cha hon ton r, nhng ngi ta xc nh c mt loi c t c cu trc khng nguyn v c ch tc ng ging vi ngoi c t ca S.shiga. Trong qu trnh gy bnh, EHEC lm tn thng xut huyt rut. EPEC: c ch cha c bit r. E. coli pht trin d dng trn cc mi trng nui cy thng thng. Mt s c th pht trin trn mi trng tng hp rt ngho cht dinh dng, hiu k kh ty tin, c th pht trin nhit t 5- 40oC. Trong nhng iu kin thch hp E. coli pht trin rt nhanh, thi gian th h ch khong 20 n 30 pht. Cy vo mi trng lng (nh canh thang) sau 34h lm c nh mi trng, sau 24h lm c u; sau hai ngy trn mt mi trng c vng mng, nhng ngy sau di y ng c th thy cn. Trn mi trng thch thng, sau 24h khun lc khong 1,5 mm, hnh thi khun lc in hnh dng S, nhng cng c th dng R hoc M. E. coli c kh nng ln men nhiu loi ng v c sinh hi. Tt c E. coli u ln men lactose v sinh hi (tr E. coli loi EIEC) .E.coli c kh nng sinh indol, khng sinh H2S, khng s dng c ngun carbon ca citrat trong mi trng Simmons, c decarboxylase (v vy c kh nng kh carboxyl ca lysin, ornitin, arginin v acid glutamic, betagalactosidase (+), VP (-). Khng nguyn O: ngi ta bit ti gn 160 yu t khng nguyn O ca E.coli. a) Chn on trc tip Bnh phm khc nhau ty bnh: l phn vi nhim khun ng tiu ha, nc tiu vi nhim khun ng tit niu, mu nu l nhim khun mu C th lm tiu bn soi trc tip i vi mt s loi bnh phm nh cn ly tm nc tiu hoc nc no ty.

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Phng php chn on ch yu nht l nui cy phn lp. Bnh phm phn c cy trn mi trng c cht c ch chn lc nh DCL, Endo. Nc tiu gia dng c tin hnh cy trn thch thng, mu c cy vo canh thang. Sau khi phn lp c vi khun thun nht th tin hnh xc nh tnh cht sinh vt ha hc v nh loi bng khng huyt thanh mu. i vi vim mng no, hin nay ngi ta cn tin hnh phng php chn on nhanh v c hiu bng k thut ngng kt latex xc nh khng nguyn ca E. coli trong dch no ty. b) Chn on gin tip Trn thc t chn on gin tip khng c s dng chn on cc nhim khun do E. coli 3.2.2. Xc nh Salmonella[11] Salmonella l trc khun Gram (-), kch thc trung bnh 3,0 x 0,5 m, c nhiu lng xung quanh thn, tr S.gallinarum v S.pullorum (gy bnh g vt). Hiu k kh ty tin, pht trin c trn cc mi trng nui cy thng thng, c th mc trn mt s mi trng c cht c ch chn lc c dng trong phn lp vi khun ny t phn. - Trn mi trng lng: sau 5-6h nui cy, vi khun lm c nh mi trng, sau 18h mi trng c u. - Trn mi trng thch thng: khun lc trn, li, bng, khng mu hoc mu trng xm. - Trn mi trng phn lp SS: khun lc c mu hng Salmonella khng ln men lactose, ln men ng Glucose thng sinh hi. S dng c citrat mi trng Simomons. Catalase (+), oxidase (-). Lysindecarboxylase (+), urease (-), RM (+), VP (-), H2S (-), indol (-). Tuy nhin khng phi bt k loi no cng c y cc tnh cht trn. Nhng ngoi l c xc nh cn phi bit l: S.typhi ln men ng glucose khng sinh hi v citrat Simmon (-). Trong loi S.paratyphi A ch c SVTH: Phm TH Hu Hu_11SHLT
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mt s chng sinh H2S v loi ny kt qu th citrat Simmon (-), lysindecarboxylase (-). S.arizona ln men lactose (tuy c chm) v onPG (-). a) Chn on trc tip Nhum soi trc tip t bnh phm Nhum soi trc tip t phn t c gi tr chn on. Thng tin hnh nhum m mt bch cu a nhn nh hng chn on. Trong bnh thng hn, mt bch cu a nhn trong phn khong 20/mt vi trng ( phng i x400). Cy mu Cy mu c tin hnh lc bnh nhn ang st cao, cn ly mu trc khi iu tr khng sinh. Ly 5 n 10 ml mu tnh mch cy vo canh thang (thng dng canh thang c mt b), 37oC. Vi khun thng hn thng mc sau 24 n 48h. Khi thy c vi khun mc, tin hnh nhum Gram, xem hnh th v tnh cht bt mu, cy chuyn sang mi trng c, quan st tnh cht khun lc, kim tra tnh cht sinh vt ha hc, cui cng xc nh tnh cng thc khng nguyn bng khng huyt thanh mu. Nu cha iu tr khng sinh, tun l u, t l cy mu dng tnh n 90%, tun th 2 khong 70- 80%; tun th 3 khong 40- 60%. Cy mu dng tnh cho php chng ta xc nh chc chn bnh nhn mc bnh thng hn. Cy phn Trong phn c rt nhiu loi vi khun, v vy, bnh phm cn c cy vo mi trng c cht c ch chn lc nh mi trng SS (Salmonella Shigella), mi trng istrati, mi trng Endo, mi trng DCLTrong nhng mi trng ny c ng lactose v cc ch th mu gip cho vic phn bit vi khun ln men v khng ln men lactose. trnh b st nhng trng hp phn c t mm bnh, trong thc t, ngoi vic cy trc tip vo mi trng c cht c ch chn lc, cn cy vo mi trng tng sinh. Sau khi vi khun mc, chn khun lc nghi ng nhum soi, xc nh tnh cht sinh vt ha hc, xc nh tnh cht khng nguyn.

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GVHD: Trn Th X

Ch ring cy phn, d phn lp c vi khun cng khng cho php ta xc nh chc chn bnh nhn mc bnh v ngi lnh cng c th mang vi khun thng hn. Cy phn ngoi mc ch chn on bnh cn c gi tr kim tra sau khi bnh nhn ht cc du hiu lm sng c cn tip tc o thi vi khun na hay khng. b) Chn on gin tip Tin hnh phn ng Widal xc inh khng th trong huyt thanh. Sau khinhim Salmonella 7- 10 ngy, trong mu s xut hin khng th O, sau 12- 14 ngy xut hin khng th H. Thi gian tn ti khng th trong mu trung bnh l 3 thng i vi khng th O v 1 n 2 nm i vi khng th H. 3.2.3. Xc nh Shigella[11] Shigella l trc khun mnh, bt mu Gram (-), di t 1-3 m, khi mi nui cy c dng cu trc khun. Shigella khng c lng, khng c v v khng sinh nha bo v vy khng c kh nng di ng. Nh cc vi khun ng rut khc, Shigella l vi khun hiu k kh ty tin nhng pht trin rt tt trong iu kin hiu kh. Trn mi trng c, chng to thnh khun lc trn, li, b u, trong v c ng knh khong 2mm sau 24h. Trn mi trng phn lp c lactose, khun lc vn khng mu. Tt c cc Shigella u ln men ng glucose, hu ht khng sinh hi, mt s trng hp c sinh hi nhng rt yu. Cc Shigella khng ln men lactose, tr S. sonnei c kh nng ln men lactose chm sau t 2 ngy n 2 tun. CcShigella flexneri, Shigella boydii v Shigella sonnei c kh nng ln men monnitol; S. dysenteriae khng c kh nng ny. Shigella khng sinh H2S, khng s dng c citrat trong mi trng Simmons, khng sinh indol. Tt c cc Shigella u c khng nguyn thn O, mt s c khng nguyn K v tt c u khng c khng nguyn H. a) Nhum soi trc tip

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Bnh phm c th ly sau khi bnh nhn i ngoi ra b sch, nn ly ch phn c nhy mu, hoc ly trc tip t trc trng, lm tiu bn nhum soi xc nh mt bch cu a nhn. bnh nhn l trc khun, mt bch cu a nhn trong phn rt cao, thng l 30 n 50, c khi trn 50 trong mt vi trng vi phng i x400. b) Cy phn Cy phn l phng php tt nht chn on bnh l trc khun. Bnh phm cn phi c cy ngay ln mi trng phn lp v vi khun l cht rt nhanh sau khi ra ngoi mi trng, nn cy trn mi trng c cht c ch chn lc nh: DCL, Endo, SS v khng c cht c ch chn lc l thch lactose- xanh bromothymol. Sau 24h, chn khun lc nghi ng, xc nh tnh cht sinh vt ha hc v nh loi bng cc khng huyt thanh mu. c) Chn on gin tip Phn ng huyt thanh rt t c lm chn on bnh l trc khun v y l bn cp tnh, cn chn on nhanh, mc khc, phn ng huyt thanh khng c tnh c hiu cao. Tuy nhin, chn on huyt thanh i khi cng c tin hnh vi nhng trng hp mn tnh, cy phn khng phn lp c vi khun, hoc lm v mc ch nghin cu dch t hc[11]. 3.2.4. Xc nh Vibrio cholerae[11] V. cholera hiu kh, nhit thch hp l 37oC, c th pht trin tt trong mi trng kim (pH 8,5- 9,5), c nng NaCl cao (3%). Trong mi trng pepton kim mc nhanh v to vng. Trn mi trng thch kim sau 18h c th quan st thy khun lc trn, li nhn v trong sut. Trn thch MacConkey khun lc trong (khng ln men ng lactose). Trn mi trng TCBS khun lc c mu vng v ln men ng sucrose. Tnh cht sinh vt ha hc Oxidase (+), indol (+), glucose (+), sucrose (+), manose (+), lactose (-), arabinose (-), H2S (-), urease (-). Kh nng khng

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GVHD: Trn Th X

V. cholerae c sc khng yu vi cc tc nhn l ha, tr pH kim. Tuy nhin c th sng 1h trong phn v mt s ngy trong nc a) Chn on trc tip Bnh phm l phn v cht nn, cn phi xt nghim ngay, nu cha c xt nghim trong vng 2h th phi cy vo mi trng bo qun CaryBlair. b) Nhum soi hoc soi ti Nhum soi quan st hnh th vi khun trc tip t bnh phm t c gi tr. Nhum soi m bch cu trong phn: trong bnh t thng thy s lng t, khong 5 bch cu trong 1 vi trng, quan st tnh di ng ca vi khun t di knh hin vi nn en. c) Nui cy phn lp Cc mi trng thng c dng phn lp vi khun t l: pepton kim, thch kim, TCBS K thut khng th hunh quang trc tip (Direct Fluorescent Antibody Test DFA) Lm tiu bn t bnh phm hoc sau khi cy vo pepton kim 6-8h. Nhum tiu bn bng khng th gn hunh quang. Ra loi b phn khng th hunh quang tha ri soi knh hin vi hunh quang. Phng php ny cho kt qu rt nhanh v c tnh c hiu cao, thng c p dng trong kim dch cc ca khu. Hin nay mt s nc sn xut b chn on nhanh, theo nguyn l k thut khng th hunh quang trc tip, s dng khng th n dng vi nhy v c hiu t 100% - Cc chng khng thuc nhm 01 cng c th gy bnh, tuy c ch gy bnh khc vi V. cholerae 01. - T nm 1992 xut hin nhm huyt thanh mi, V. cholerae 0139, gy ra nhiu v dch t nhiu nc trn th gii- Cc V. cholerae khng thuc nhm 01 ch khng ngng kt vi khng huyt thanh 01, chng s b ngng kt khi gp khng huyt thanh c hiu. Thu thp bnh phm SVTH: Phm TH Hu Hu_11SHLT
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GVHD: Trn Th X

Mu phn nn thu thp sm, trc khi iu tr khng sinh. Mu phn cha c lm ngay phi c bo qun mi trng vn chuyn Cary-Blair, vi khun t c th sng 4 tun trong mi trng ny. C th s dng mi trng canh thang giu cht dinh dng lm mi trng vn chuyn: nc peptn kim v canh thang peptn tellurite-taurocholate. Gi lu bnh phm nc peptn kim nhit phng, cc loi Pseudomonas v vi khun t 01 v non-01 s pht trin ln t vi khun t 01. Cc mu bnh phm gi trong mi trng vn chuyn nn gi ti phng th nghim, khng trong t lnh v vi khun t rt nhy cm t lnh. Soi trc tip Lam knh 1: Ho mu phn vo nc mui sinh l, nh 1 git ln lam knh, y phin knh mng (lamen) ln trn. Soi tiu bn trc tip trn knh hin vi thy vi khun t di ng rt nhanh theo nhng ng thng t u n cui vi trng. Lam knh 2: Nh 1 git huyn dch phn ln lam knh, nh 1 git khng huyt thanh t a gi. Nu l vi khun t th vi khun s b bt hot, khng di ng na. Soi tiu bn trn knh hin vi nn en, vi khun t di ng nh sao i ngi. Vi khun t l trc khun Gram m, hnh cong du phy. Mi trng tng sinh: pepton kim mn 2 - 3 ln/ 6 - 8 gi/ 370C. Mi trng nui cy: Trn mi trng thch kim: khun lc t trn nh, trong, t, long lanh nh ht sng. Trn mi trng TCBS (Thiosulphate Citrate - Bile salts - Sucrose): khun lc t mu vng, to v nhn. Chn khun lc nghi ng t a mi trng phn lp, cy chuyn sang cc mi trng xc nh tnh cht sinh vt ho hc[13]. 3.2.5. Xc nh Shigella[11] Shigella l ging vi khun nguy him, thuc h vi khun ng rut Enterobacteriaceae, gm 4 loi ph bin S.flexerneri, S.dysenteriae, S.sonnei v S. boydii, gy bnh l cho ngi khi hin din mt t 10 n 20 t bo. SVTH: Phm TH Hu Hu_11SHLT
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Bnh do nhim Shigella c th gy ra hi chng Reiter, hi chng HUS v dn n t vong cao (t l 0.7%). Gen m ha cho c tnh ca vi khun Shigella c th nm trn nhim sc th hay trn plasmid xm ln. Nhng gen ny thng chu tc ng iu ha a yu t v gen c khuynh hng tp trung thnh cm trong b gene. Cc phng php PCR pht hin cc gen c tnh gy bnh ca Shigella thng tp trung vo 3 gene: gene virA (nm trn NST), ipaH v ial (2 gene nm trn plasmid). Gene ial thng c tm thy trn plasmid ln c khuynh hng b mt i trong qu trnh di truyn, c th gy ra hin tng m tnh gi khi pht hin gene ny. Gene ipaH hin din trn c NST v plasmid ca Shigella, v vy l mt gene c tnh n nh cao hn pht hin. Kit nh tnh Shigella da trn k thut PCR in di pht hin s hin din vi khun Shigella trong mu thc phm. Quy trnh PCR s dng mt cp mi nhn bn mt phn on gene iapH, c chng minh hin din rng ri cc chng Shigella trong nhiu nghin cu trn th gii v Vit Nam. S hin din sn phm PCR kch thc 423bp trn bn gel agarose cho php kt lun mu nhim khun Shigella. Quy trnh thc hin gm 4 bc (1) Tng sinh trn mi trng Buffered Peptone Water (2) Tch chit, thu nhn DNA (3) Nhn bn DNA bng PCR (4) in di xem kt qu. Ton b quy trnh thc hin trong khong 30 gi

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KT LUN Tnh hnh ng c thc phm v nguy c nhim thc phm ang l mt trong nhng thch thc trong vic bo m v sinh an ton thc phm hin nay. Ng c thc phm v ang e da trc tip n sc khe, tnh mng ngi tiu dng, nht l cng nhn ang lm vic, sinh hot ti cc nh my, khu cng nghip, nh hng n mi hot ng kinh t, an sinh x hi. Phng, chng ng c thc phm hiu qu, bn vng ang i hi s n lc v s hp tc cht ch, thng xuyn ca cc cp, cc ngnh, cc a phng v ca c cng ng. Thng qua n ny em gip cho em hiu bit nhiu hn v tnh hnh ng c thc khng nhng th n cn bi p cho em nhng kin thc chuyn ngnh cng ng sinh. Gip cho em c thc hn v vn gi gn v sinh an ton thc phm v c th vn ng mi ngi xung quanh phng trnh nhng ri ro do thc phm khng an ton gy ra.

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PH LC

Hnh 3.2. E.coli, Salmonella, Shigella

Hnh 3.3. Phn lp E.coli trn MacConkey - Phn lp E.coli trn CHRO Magar SVTH: Phm TH Hu Hu_11SHLT
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Hnh 3.4. E.coli di knh hin vi in t

Hnh 3.5. Salmonella pht trin trn thch mu

Hnh 3.6.Salmonella trn thch McConkey-Salmonella sau 24h trn thch XLD

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Hnh 3.7. Shigella trn thch HE, khun lc trn, b u v trong

Hnh 3.8. Nui cy Shigella trn thch McConKey

Hnh 3.9. Shigella di knh hin vi

Hnh 3.10. Vibrio cholerae di knh hin vi SVTH: Phm TH Hu Hu_11SHLT

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GVHD: Trn Th X

Hnh 3.11.Phn lp Vibrio trn MacConkey- Phn lp Vibrio trn CHROMagar TI LIU THAM KHO [1].http://tcyh.yds.edu.vn/96-02/1996-1997/thuc%20an%20&%20cac %20benh%20nhiem%20trung%20tieu%20hoa.htm [2].http://www.yduocngaynay.com/88TK_NgTChanh_NgoDocThucPham.htm [3].http://vi.wikipedia.org/wiki/Ng%E1%BB%99_%C4%91%E1%BB %99c_th%E1%BB%B1c_ph%E1%BA%A9m#Vi_khu.E1.BA.A9n [4].http://vietbao.vn/Suc-khoe/8-trieu-nguoi-ngo-doc-thuc-pham-moinam/30189842/248/ [5]. http://www.t5g.org.vn/Default.aspx?u=cmdt&grnid=102&cmid=24 [6].http://www.victoriavn.com/index.php/comp:news/id:397/Cac-loai-vikhuan-thuong-gay-ngo-doc-thuc-pham.html/# [7].http://kiemnghiemthucpham.blogspot.com/2011/01/28-tcn199-2004salmonella-trong-san.html [8]. http://bancuabe.org/index.php? mod=entries&act=detail&yt=2&vd=2&id=20 [9].http://ngochungtihe.blogspot.com/2011/01/xac-inh-cac-loai-vibriotrong-thuc-pham.html SVTH: Phm TH Hu Hu_11SHLT
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[10]. JEJUNI

GVHD: Trn Th X

http://www.scribd.com/doc/65381143/19/I-CAMPYLOBACTER-

[11].http://www.impe-qn.org.vn/impe-qn/vn/portal/InfoDetail.jsp? area=58&cat=1101&ID=2761 [12]. http://www.scribd.com/doc/31588247/Tieu-Luan-Salmonella [13]. http://www.scribd.com/doc/55372088/BAO-CAOSTAPHYLOCOCCUS-2 [14].http://kt-biotech.com/vi/kit-chan-doan-vi-khuan-listeriamonocytogenes/39-kit-lm-dinh-tinh.html [15].http://blogthuysan.blogspot.com/2011/10/cach-kiem-tra-vi-sinh-vathieu-khi.html

MC LC

SVTH: Phm TH Hu Hu_11SHLT

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