Ky Sinh Trung Thuc Hanh

B Y T
GIO TRNH

K SINH TRNG
THC HNH
(DNG CHO O TO C NHN XT NGHIM)
M S: K.01.Z.15

NH XUT BN GIO DC
H NI 2008

Ch o bin son:
V KHOA HC V O TO B Y T

Page 1 of 218 LI GII THIU
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Ch bin:
PGS. TS. L TH XUN

Nhng ngi bin son:
CN. V TH M DUNG
CN. NGUYN TH HIN
CN. TRNH TUYT HU
CN. NGUYN H PHNG LIN
PGS.TS. L TH XUN

Tham gia t chc bn tho:
ThS. PH VN THM
TS. NGUYN MNH PHA

LI GII THIU

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 C nhn xt nghim. 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.
Gio trnh K SINH TRNG THC HNH c bin son da vo chng trnh gio dc ca Trng
i hc Y Dc TP H Ch Minh trn c s chng trnh khung c ph duyt. Gio trnh c
PGS.TS. L Th Xun (Ch bin), CN. V Th M Dung, CN. Nguyn Th Hin, CN. Trnh Tuyt Hu, CN.
Nguyn H Phng Lin 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.
Gio trnh K SINH TRNG THC HNH c Hi ng chuyn mn thm nh sch v ti liu
dy hc chuyn ngnh C nhn xt nghim 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 v Hi ng chuyn mn thm nh gip hon thnh cun
sch; cm n PGS.TS. V c Chnh, PGS.TS. Hong Tn Dn 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

Gio trnh K sinh trng thc hnh c bin son cho sinh vin khoa K thut Y hc c mc ch
hng dn cho nhng sinh vin hc mn K sinh trng nhm hon thin kh nng chn on da trn mt s
thng tin lm sng c bn v xt nghim bnh phm bng cch xem knh hin vi, cy. Mt s k thut min
dch cng c cp n.
Gio trnh gm c ba phn:
Phn mt: Phn k thut trnh by nhng k thut xt nghim c bn bao gm phng php thu thp,
bo qun, x l bnh phm.
Phn hai: nh danh, gm cc hnh nh cc k sinh trng v vi nm gy bnh thng gp nc ta.
Ngoi vic cc sinh vin phi nm vng cc k thut c gii thiu, iu chng ti quan tm hn na
l sinh vin phi bit c u, nhc im ca cc phng php c chn, phi hiu ch li v hn ch ca
n. Sinh vin cn phi bit la chn cc phng php chn on ph hp vi tng loi k sinh trng v tng
loi bnh phm.
Ni dung cc k thut trnh by trong gio trnh ny c th khng c y , nhng n cng cha
ng cc phng php ph bin nht v dng cho cc phng xt nghim lm sng nc ta.
Trong cun gio trnh ny, chng ti c gng trnh by nhng im c trng v hnh th phn
bit k sinh trng v gii thch lm th no xc nh chng.
Phn ba: Ph lc, gii thiu cc ha cht thng dng trong xt nghim k sinh trng ng rut; cc
ha cht, thuc nhum v mi trng trong xt nghim nm.
Nhng hnh nh minh ha, mc d khng hon chnh nhng cng kh y v s lng v cht lng,
cung cp mt cch khi qut v hnh thi ca k sinh trng v vi nm cng nh cc k thut pht hin chng.
Cc tc gi l nhng ngi lm vic phng th nghim trong nhiu nm qua v c kinh nghim ging
dy v mn K sinh trng, hy vng rng cun sch ny s cung cp nhng thng tin c gi tr cho sinh vin
nhm gip h c kin thc v thc tin chn on k sinh trng, gip cho vic phng, cha bnh t hiu
qu.
Do trnh v thi gian c hn, chng ti khng trnh khi nhng thiu st v chuyn mn cng nh in
n, rt mong nhn c s gp ca cc sinh vin v ng nghip ln xut bn sau gio trnh c hon
thin hn.
Xin chn thnh cm n.

CC TC GI

PHN MT
K THUT

Bi 1
CCH S DNG V BO QUN KNH HIN VI


a s k sinh trng (KST) khng th nhn thy bng mt thng m cn c nhng dng c quang hc
phng i chng ln nh knh lp, knh hin vi. Ty theo yu cu ca k thut, knh hin vi cn cn c
nhng ph tng o kch thc KST, t quang nn en,
1. NHC LI CU TRC CA KNH HIN VI
Knh hin vi l mt cng c thng dng v quan trng nht ca mt phng xt nghim KST. Knh hin
vi c th c nhng hnh dng khc nhau ty theo mu sn xut, nhng cu to c bn ging nhau, gm c
nhng b phn:
C Th knh l mt thu knh nm pha trn mt nhn nh qua vt knh. C 3 loi th knh x5, x10,
x15; loi x10 thng c dng nhiu nht.
C ng knh l mt ng m nh sng phi i qua t vt knh n th knh v c chc nng gi th knh v
vt knh nm cch nhau mt khong nht nh.
C a mang vt knh l mt b phn c 4 l gn vt knh, khi xoay s a vt knh cn s dng vo
ng knh.
C Vt knh: nh sng i qua vt quan st ri n thu knh ny. C 4 loi vt knh, nhng thng dng 3
loi:
Vt knh x10: c th trng ln nht, sau khi iu chnh thy r mu vt, vt knh ny thng cch
knh mang vt khong 16mm.
Vt knh x 40: c phng i trung bnh, sau khi iu chnh thy r mu vt, vt knh ny thng
cch knh mang vt khong 4mm.
Vt knh x100: c phng i ln nht, sau khi iu chnh thy r mu vt, vt knh ny thng
cch knh mang vt khong 1mm. S dng vt knh vi du soi knh v dng c vi cp iu chnh.
C Knh t quang: tp trung nh sng.
Mng chn nh sng: cho nh sng qua nhiu hay t vo vt knh.
C Gng trn dng ly nh sng, thng c 2 mt:
Mt lm: khi s dng vt knh x10, x40.
Mt phng: khi s dng vt knh x100.
Nhng loi knh dng nh sng ca bng n gn trong thn my khng c gng.
Tiu xa: dng gi tiu bn c gn vi mt trc c mt c dng di chuyn sang tri, sang phi
v mt c dng di chuyn pha trc, v sau.
Thn knh mang ng knh, bn mang mu vt, knh t quang, c vi cp, c th cp v gng.
1 Chn: c chc nng gi cho knh c vng v n nh.

Cu to knh hin vi quang hc
2. CCH S DNG KNH HIN VI
C t tiu bn ln bn mang tiu bn.
C iu chnh nh sng vi gng trn, knh t quang v mn chn sng.
C Xoay trc mang vt knh x10 vo ng v tr.
C Vn c th cp thy r vt.
C Nu cn quan st vi phng i ln th i qua vt knh ln hn x40, dng c vi cp iu chnh
n khi thy r vt. Khi s dng vt knh x100, ta phi dng du soi knh. Nh 1 git du ln tiu bn ri i
qua vt knh x100.
3. CCH BO QUN KNH HIN VI
C t knh hin vi ng ch, xa hi nng v ch m t.
C Cm knh hin vi bng thn knh, tay kia chn ca knh. Phi ng knh hin vi, khng c
knh nghing.
C Cn thn khng lm ri cht n mn hay bt c mt dung dch no ln bn knh.
C Khng c tay t hay bn ln knh hin vi.
C Lau th knh v vt knh bng giy lau knh trc v sau khi dng. Khi soi vi vt knh du, thm giy
lau knh vi mt git xylen lau vt knh. Sau khi lau vi xylen, phi lau kh ngay bng giy lau knh, nu
khng xylen c th lm bong nhng thu knh gn trong vt knh.
Trc khi ct knh hin vi, vt knh nh v tr quan st v h thp ng knh bng c ln. Vn nh
nhng, ng n mnh ng knh. Nu cn thn hn, h t quang knh xung. Nu t quang knh bn, lau bng
giy lau knh kh.
C gng nghing, mt phng ra pha ngoi trnh bi.
Che knh hin vi bng bao ca knh. Ct knh vo ng ch ca knh, lui vo pha trong, ng
mp m pha ngoi.

CU HI LNG GI
1. Trnh by cch s dng knh hin vi quan st mt mu phn ti.
2. Khi s dng knh hin vi soi lam mu, anh (ch) cn ch n yu t no c th nhn thy r KST
st rt (KST SR) trn pht mu nhum?
3. Sau khi soi lam mu tm KST SR, anh (ch) bo qun knh hin vi nh th no trc khi ct vo t knh?

Bi 2
CCH CHUN KNH HIN VI

Xc nh loi KST cn da vo nhiu tiu chun, trong c tiu chun kch thc ca KST. Ta c th
c lng kch thc KST bng cch so snh vi mt vt bit kch thc trc nh hng cu, nhng cch
ny khng cho ta kt qu chnh xc. o c chnh xc kch thc KST, ta dng thc trc vi t trong
th knh.
1. DNG C
Knh hin vi 2 mt vi cc vt knh x 10, x 40, x 100
Du
Giy lau knh
Thc trc vi th knh (chia thnh 50 n v)
Thc trc vi nn vi 2 chia 0,1 v 0,01mm
Th knh (nn s dng th knh x10):
+ Thc trc vi nn c kch thc bng lam knh bnh thng v gia c nhng gch cch nhau 0,1 v
0,01mm
+ Thc trc vi t th knh l mt ng thng c chia thnh 50 vch. Ty theo phng i ca
vt knh, cc vch ny c cc s o khc nhau.

2. QUY TRNH K THUT
C Tho th knh ra v t thc trc vi th knh vo (mt khc vch hng xung di). t th knh tr
li v tr c.
C t thc trc vi nn ln bn knh hin vi.
C Di chuyn bn knh sao cho 2 thc nm chng ln nhau, vch 0 trn thc trc vi th knh trng vi
vch 0 trn thc trc vi nn.
C Nhn pha bn phi vch 0 ca thc trc vi nn tm im m 1 vch ca thc trc vi th knh
trng vi 1 vch ca thc trc vi nn, im trng ny gi l im Y. Khong cch s thay i ty theo cc
vt knh s dng (x10, x40, x100).
C m s vch chia trn thc trc vi th knh, t s 0 n vch trng lp (Y). m s vch chia
(0,1mm) trn thc trc vi nn t vch 0 n vch trng lp (Y), Tnh on m c trn thc trc vi th
knh theo cng thc sau:

N = S vch m c trn thc trc vi nn (mm).
n = S vch m c trn thc trc vi th knh (mm).
V d: vt knh x10, ta c N = 0,3mm, n = 40.

V d: o chiu di ca trng giun kim.
t tiu bn ln bn knh, quan st trng vi vt knh 10, chiu di ca trng giun kim tng ng vi 8
khong chia ca thc trc vi th knh.
Ta c n v th knh vt knh x10 l 7,5mm, chiu di ca trng giun kim s l 7,5mm x 8 =
60mm.
Lu :
Mi phng i ca vt knh (x10, x40 v x100) c n v th knh khc nhau, v mi vch ca
thc trc vi nn s thay i kch thc trong khi vch ca thc trc vi th knh vn duy tr kch thc c.
V vy, cn phi chun cho tng loi vt knh v ghi li cc n v ny ln knh hoc t giy dn gn knh
d tra cu.
Khi mun c s o ca KST th ch cn nhn s vch o c vi n v th knh c kch thc
tht.
Sau khi mi vt knh c chun , ta khng trao i th knh cha thc trc vi v nhng vt
knh ca knh hin vi ny vi th knh hoc vt knh ca knh hin vi khc. Phi s dng vt knh v th knh
c chun .
Nn chun nh k bo m tnh chnh xc.

CU HI LNG GI
1. To sao cn phi bit kch thc ca KST?
2. Trnh by cch tnh n v th knh.
3. Lm th no o kch thc ca trng giun a?

Bi 3
THU THP V BO QUN PHN
XT NGHIM TM K SINH TRNG

1. THU THP BNH PHM
C nhiu phng php ly bnh phm, vic quyt nh chn phng php no da vo gi tr v gii
hn ca mi phng php. Nu bnh phm khng c ly v x l ng yu cu k thut, chng ta c th
khng pht hin c mm bnh.
1.1. Chun b bnh nhn trc khi ly phn
Nhiu kt qu xt nghim phn l m gi to do bnh nhn khng c hng dn y hay hng
dn khng ng cch. Phi hng dn bnh nhn mt cch cn thn; tt nht l phng th nghim a cho
bc s iu tr nhng bn in sn nhng chi tit cn thit pht cho bnh nhn c ch nh xt nghim
phn.
Dn bnh nhn trong 3 ngy trc khi ly bnh phm, trnh dng nhng loi thuc v thc phm c th
lm cho vic nhn dng KST kh khn nh:
Thuc: Bismuth, Magnesium, Kaolin, Baryte, thuc t vo hu mn c du, m.
Thc phm nhiu cn b: ng cc, bp ci, salad, qu c nhiu ht nh, nhiu cht bo, du, m.
Bnh nhn nn n ch t cht b nh: bnh, n long, trng, sa, gan,.
1.2. Ly bnh phm
1.2.1. Ti phng xt nghim
Tt nht nn ly phn ti phng xt nghim.
L ng phn:
+ Cn phi kh v sch, bng nha trong hoc giy carton khng thm nc hoc thy tinh.
+ C ming rng, np vn cht.
+ C dn nhn ghi h, tn, tui, a ch ca bnh nhn v ghi ngy, gi ly bnh phm.
Cch ly phn:
+ C th ly bt c ch no ca khun phn tm trng giun, sn. Nhng pht hin n bo, nn ly
phn ch bt thng nh mu, nhy, lng, bt hoc ly phn ngay trong trc trng.
+ Khng c ly phn ln vi nc tiu, du, cc cht mui Mg, Al, Ba, Bi, Fe v cc cht lm bin
dng n bo.
+ Nu cho bnh nhn ung thuc x, ch nn cho ung sulfat natri v s ly phn khi bnh nhn i ngoi
ln th hai hay th ba sau khi ung thuc.
Lng phn cn ly:
+ Thay i ty theo mc ch v k thut xt nghim, thng ch cn khong 5 10 gam phn (khong
bng ht lc) c th lm nhiu phng php.
+ Trong mt s trng hp nh tm giun, t sn, cc bnh v b tiu ho phi ly ton b s lng
phn c thi ra c th thy c KST v mng nhy hay m b b trc ra cng vi phn.
1.2.2. Ngoi phng xt nghim
Ly phn ngoi phng xt nghim l iu bt c d, cn tn trng nhng nguyn tc sau:
Phi gi n phng xt nghim trong thi gian ngn nht, c bit l n bo, phn phi lun c gi
m.
Khng c gi nhit lnh qu.
Nu xa: gi hp phn trong nc m 37
o
C v ng thi ly mt cht phn cho vo mt trong nhng
dung dch c nh:
+ MIF: Merthiolate Iod Formol.
PVA: Polyvinyl Alcohol.
F2AM: Formol + Phenol + Alcool + Xanh Methylene.
1.3. Thi gian xt nghim phn
Sau khi thu hi bnh phm cn xt nghim ngay, cng sm cng tt. Thi gian t khi ly mu n khi
kho st:
Phn bnh thng cn xt nghim trong vng 12 24 gi hoc c th 1 2 ngy trong t lnh.
Phn mm, nho, lng hay c mng nhy v mu cn phi xem ngay trong vng 30 pht sau khi ly.
Trong trng hp sau khi ly phn m cha xt nghim ngay hoc ly phn ti nh xa, nn bo qun
phn bng cch phn trong cc dung dch nh hnh (fixative) trng giun, sn khng pht trin, n bo
khng b thoi ha.
2. HA CHT BO QUN PHN
bo qun hnh th v ngn s pht trin tip tc ca trng v u trng giun, sn, phn c ng
trong cht bo qun ngay lp tc sau khi ly (bnh nhn ly) hoc khi phng xt nghim nhn bnh phm.
Mt s cht c nh c a dng l: formol, sodium acetatacetic acidformol (SAF), dung dch
Schaudinn, polyvinyl alcohol (PVA).
Khi chn phng php c nh, phi m bo cht c nh c chn ph hp vi k thut xt nghim
s lm. V mi cht c nh c tnh cht ring, khng th dng cho tt c cc loi k thut xt nghim.
2.1. Formol
Formol c bit thch hp c nh u trng giun, sn v bo nang n bo. Hai nng thng dng
l 5% cho bo nang n bo v 10% cho trng v u trng giun, sn.
gi hnh dng n bo c tt, nn pha long formol vi dung dch m phosphat, to thnh formol
trung ha.

Ghi ch: Formaldehyd bn th trng thng ch 37 40% HCHO, tuy nhin vn c xem l 100%.
Bo nang n bo, trng nang ca trng bo t, trng giun, sn v u trng c bo qun lu di trong
formol 10%. Formol nng (60
O
C) c th dng i vi bnh phm c trng giun, sn (v trong formol lnh,
mt vi loi trng dy s tip tc pht trin, gy nhim v sng trong mt thi gian di).
Ly vi gram phn trn k trong dung dch formol 5 10%.
u im:
C nh ton b phn.
Pha ch d, bo qun lu.
Cn lng c th lm th nghim min dch.
Nhc im:
Khng bo qun th hot ng.
Hnh dng KST khng p trn pht nhum c nh.
2.2. Sodium acetat acetic acid formol (SAF)
SAF c dng bo qun trng v u trng giun, sn, bo nang v th hot ng n bo, trng nang
trng bo t v bo t Microsporidia.
Bnh phm c nh trong SAF u dng c vi phng php tp trung phn v lm pht nhum c
nh. Khi lm pht phn nhum, nn trn thm albumin vo phn tng dnh ca bnh phm vo lam
knh.
SAF c coi l cht c nh mm hn thy ngn clorua. Hnh dng KST s khng sc nt bng khi c
nh trong dung dch c thy ngn clorua. Kt hp c nh SAF vi nhum hematoxylin st cho hnh dng
tt hn nhum Trichrome.
Thnh phn:

Pha ch Albumin Mayer: trn mt th tch lng trng trng vi mt th tch glycerin. Cho mt git
hn hp ny ln lam knh, cho thm mt git cn lng phn SAF, trn u, kh nhit phng 30 pht
ri nhum.
u im:
Dng cho tiu bn tp trung v c nh.
Khng c hp cht thy ngn.
D pha ch, bo qun lu.
Cn lng c th lm k thut min dch men.
Nhc im:
Bnh phm t bm vo lam knh.
2.3. Dung dch Schaudinn
c dng vi phn ti hoc bnh phm nim mc rut, c th dng cho tiu bn nhum c nh v
phng php tp trung.
Cch pha ch:
Dung dch thy ngn clorua bo ho:

Dng mt cc chng, un si n khi thy ngn clorua tan. yn vi gi n khi to tinh th.
Dung dch c nh Schaudinn (dung dch m):

Thm 5ml acid acetic lnh vo 100ml dung dch m ngay khi s dng.
u im:
C nh tiu bn t mu phn ti hoc nim mc rut.
Bo qun tt th hot ng v bo nang n bo.
Nhc im:
Khng khuyn co dng trong phng php tp trung.
Dung dch c cha thy ngn clorua, nn t ra vn x l nc thi.
t dnh vi bnh phm lng hoc nhy.
2.4. Polyvinyl alcohol (PVA)
PVA l mt nha do phi hp vi dung dch c nh Schaudinn. Bt PVA c dng nh cht dnh
cho bnh phm phn khi hn hp phn PVA c tri trn lam knh, cn vic c nh vn l dung dch
Schaudinn.
Dung dch c nh PVA c dng bo qun tt c cc th ca KST ng rut, nht l bo nang v
th hot ng n bo cho nhng k thut chuyn su.
PVA cng c dng c nh bnh phm cn gi qua bu in n nhng phng th nghim chuyn
su, rt tt vi bnh phm lng v pha theo t l 3 phn PVA vi 1 phn phn.
Cch pha ch:

Trn cc dch lng vo cc 500ml, thm bt PVA vo (khng khuy). y cc bng a Petri ln, giy
sp hoc l kim loi, qua m. un t t n 75
o
C, ly cc ra v khuy trong khong 30 pht n khi c
dung dch ng nht nh sa.
u im:
C th lm tiu bn c nh v phng php tp trung.
Bo qun tt bo nang v th hot ng n bo.
Bo qun lu (hng nm) trong l kn nhit phng.
Bnh phm c th gi bng bu in n phng th nghim chuyn su.
Trng Trichuris trichura v bo nang Giardia lambia trong phng php tp trung d nhn ra nh
trong phng php formolether.
Nhc im:
Hnh dng u trng Strongyloides stercoralis khng p nh c nh bng formol. Trng nang
Isospora belli c th khng quan st c (formol tt hn).
Dung dch c cha thy ngn, nn t ra vn x l nc thi.
C th tr nn trng v st do mt nc hay do lm lnh.
Kh pha ch trong phng th nghim.
Khng th dng tiu bn lm k thut min dch men.
2.5. PVA ci tin
PVA c ci tin khng cha thy ngn, m thay vo ngi ta dng sulfat ng hoc sulfat km.
Sulfat ng khng cho kt qu tt nh thy ngn clorua. Sulfat km c dng trong nhum Trichrome.
u im:
Dng c cho pht nhum c nh v phng php tp trung.
Khng cha thy ngn.
C nh bng sulfat km cho kt qu tt hn v th nhiu ngi thch dng PVA c cha sulfat km
hn sulfat ng.
Nhc im:
Hnh dng ca bo nang v th hot ng n bo kh thy khi c nh bng sulfat ng, c bit khi
so snh vi thy ngn clorua.
c im cu to ca n bo khi nhum khng n nh: c th r, c th khng r. V vy, vic nh
danh c th gp kh khn, c bit i vi nhng bo nang ca n bo nh nh Endolimax nana.
3. K THUT LU GI K SINH TRNG TRONG BNH PHM
Gi bnh phm 40
o
C, c th gi c trng giun, sn v bo nang n bo nhiu ngy, nhiu tun m
vn c th nh danh c d dng. Mun gi lu phi dng dung dch nh hnh.
3.1. Trng, u trng giun, sn v bo nang n bo
a) Lu gi trn tiu bn lm t phn t
trnh loang phi dng lam knh sch, ra sch m trong dung dch ng th tch cn ether.
trnh kh do tip xc vi khng kh, khng bay hi, ngi ta hn tiu bn bng:
+ Vaselin: mu khng gi lu hn vi gi, dng quan st KST sng.
+ Paraffine:
Loi paraffine dng cho m hc, h nng chy hay t m 56
o
C. Phng php ny d thc hin
nhng tiu bn d b v khi va chm.
+ Thuc sn mng tay:
* Phi tiu bn bc hi, kh bt vin hn, n xem cn h khng. Sau hn ln th 2 v kh.
Phng php ny gin d khng cn dng c c bit.
* Nu hn kn, mu lu gi tt, trng giun, sn cn nguyn vn; ngc li bo nang v dng hot ng
lu gi khng tt.
b) Lu gi KST lu di
Dng dung dch formol m nng ty thuc vo cng ca phn (phn rn dng formol 5%; phn
st: formol 10%), phn c trng giun c sc chu ng cao (20%, F2AM).
Quy trnh thc hin:
C Cho phn vo dung dch c nh theo th tch:
1 th tch phn + 3 th tch dung dch bo qun.
C Nghin u, lc qua li kim loi loi nhng cn b ln.
C 1 pht bnh thy tinh c chn loi tr nhng phn t nng.
C phn trn vo chai c nt v c nhn.
C Ghi KST c trong mu, ngy ly mu, nng dung dch c nh.
+ KST c gi tt cn lng trong chai.
+ lm gim s bc hi ca formol, thm vo dung dch bo qun 10% glycerine.
i vi dng hot ng ca amp.
+ Dng dung dch c nh v lu gi k trn, dung dch formol 10% ch gi c amp vi tun, sau
amp s b ly gii.
+ Dung dch MIF: dung dch ny t tin nhng gi c dng hot ng ca amp nhiu nm.
+ Dung dch PVA: gi c dng hot ng ca amp v c th lm pht nhum Hematoxyline st.
i vi dng hot ng ca trng roi ng rut:
Nhng cch k trn u dng c nhng khng tt v nhng dng hot ng thng thu trn li, khng
thy c nh khi quan st trc tip. Dung dch tng i tt l MIF, F2AM.
3.2. Giun, sn trng thnh
a) Giun, sn tm thy cht trong phn
t c gi tr v chng thng b hy hoi.
Ha cht thng dng l formol 5% hoc cn ethylic 70
0
.

b) Giun, sn cn sng trong phn
Ra bng nc mui sinh l.
Phng thc c nh thay i ty theo loi giun, sn:
+ Giun:
* Ly giun ra khi nc ra trong hp Petri hay bt s.
* ngay cn ethylic 70
0
si (un si cn trong bnh Erlenmeyer c khuy t). Cch c nh ny
lm gin giun ngay lp tc.
* Gi trong bnh thy tinh c nt mi. Khng y bng nt bc hay cao su v s lm h mu mau
chng.
Lu :
* Khng c nh bng cn lnh.
* Khng lm cht giun trong NaCl 0,85%.
* Khng dng dung dch formol.
+ Sn dy v sn l:
* Kp sn gia 2 lam knh trng thi tri rng, c trong hp Petri ln.
* Cho dung dch c nh:
Cn ethylic 70
0
si.

Dung dch ng th tch dung dch formol 10% v cn 70
0
si. Ngm sn ti thiu na gi ri mi
ly sn ra.
Lu mu:
* B dung dch c nh.
* Lu mu trong cn 70
0
, trong chai thy tinh nt mi hoc nt cao su.

c) Loi giun c kch thc nh
tng l giun trong ng nghim cha cn ethylic 70
0
, y nt bng gn khng thm nc, bao
ming.
Phi dn nhn, vit ngy bng bt mc tu, bt m.
c l vo bnh c np, di lt bng thm nc, y cn ethylic 70
0
, y np. Trnh cn bay
hi (np phi c vng cao su).
3.3. Nhng iu cn bit khi lu gi KST lu di
Giun mt trong v mu t nhin: khi b c nh tr nn c v hi trng nhng gi c rt lu
trong cn.
Trng giun, sn th d nhn nhng khng ging ht nh trong phn ti. Vi loi trng nh trng giun
a, trng giun mc s b phn bo nu dung dch c nh khng nng (dng phn bo khng bao gi
gp trong phn ti).
Bo nang n bo dng ti th nhn c mu km. Sau mt thi gian lu, nhng nhn ny khng
nhum mu nhng li r hn l trng thi ti.
Dng hot ng ca amp mt nhanh chng chit quang trong dung dch formol.
Trong MIF, amp khng b ly gii, nhn ra d. Ngc li, sau khi trn lam knh v y bng l knh,
mu ca chng bin mt, khng th dng lm mu lu di trn lam knh v l knh.

CU HI LNG GI
1. Th no l thu thp phn ng quy cch?
2. i vi anh (ch), iu g quan trng trong khu thu thp phn?
3. Theo anh (ch) hin nay, cc phng xt nghim trong nc ta c ch trng n vic ly bnh phm? v
kt qu ca vic c/khng ch trng n vic ly bnh phm l g?
4. Bo qun bnh phm c ch li g?
5. Nu tn nhng ha cht bo qun phn thng c dng, cho bit u v nhc im ca tng ha cht
bo qun c nu.
6. Cch bo qun n bo khc vi cch bo qun giun, sn nh th no?
7. Ha cht bo qun c nh hng g n KST khi KST c ngm trong thi gian lu di?

Bi 4
K THUT XT NGHIM PHN TM K SINH TRNG


I. I CNG
Trong phng xt nghim, khi nhn c bnh phm, nu l phn ti khng c cht bo qun, chng ta
cn quan st bng mt (i th) trc c c nhng nhn xt s b v mu phn, ghi nhn nhng c
tnh ca mu phn, phn loi bnh phm xt nghim: mu phn lng, c cht nhy, mu phi xt nghim
ngay.
Khng nn phn ngoi tri, khng c np y; khng nn l phn trn phiu xt nghim hoc
dn mu vo cui bui mi xt nghim.
i vi phn c bo qun trong dung dch c nh th quan st i th khng thc hin c.
pht hin c KST chng ta cn phi dng knh hin vi quan st (vi th).
1. QUAN ST I TH
Quan st i th (bng mt hoc knh lp) ghi nhn trng thi, mu sc, cc cht l, tm kim v xc
nh cc loi giun, sn c thi ra theo phn.
1.1. Trng thi phn
Phn c th cc trng thi:
Cng rn (kh m thng).
Cng (m thng c).
Mm (ct c).
Nho (c th bin dng).
Lng.
Lng nh nc.
1.2. Mu sc
Thay i t en, nu m, nu, nu nht, vng, xanh, mu t st hay i khi , trng.
1.3. Cc cht l
Cht nhy: thng c, c th kt thnh si, hnh dng ging nh k sinh trng. Cht ny c xem
xt cn thn tm cc n bo, cc trng Schistosoma.
M lin kt: mu trng nh x c. Xem di knh hin vi sau khi lm trong vi acid acetic s thy
nhng si di.
Mu: ch cn ghi nhn s hin din mu ti hoc c tiu ha lm phn c mu en u.
M: gm c nhiu bch cu bin dng.
Cc cn b thc vt cha tiu ha, thng di hnh thc si.
2. QUAN ST VI TH
Quan st vi th c th c thc hin vi k thut xt nghim phn trc tip, tp trung KST trong phn,
k thut chuyn bit, cy v nhum c nh.
II. K THUT XT NGHIM PHN TRC TIP
K thut xt nghim phn trc tip s dng phn ha tan trong nc mui cho php pht hin s di
ng ca th hot ng n bo, trng giun, sn, u trng giun v cc vt th bt thng trong phn (hng
cu, bch cu,).

1. DNG C
Knh hin vi
Lam knh, l knh
Vit (bt) ch sp
Que g
Khn vi
Bnh ng dung dch st trng
Kp.
2. HA CHT

3. QUY TRNH LM TIU BN PHN
C Ly mt tm lam knh sch, kh. Dng vit ch sp chia lam knh ra lm 3 phn. Ghi tn bnh nhn
vo nh u lam knh.
C Nh ln lam knh 1 git NaCl 0,85% vo gia, 1 git Lugol cui.
C Dng que g ly mt t phn bng u que dim, ha tan phn vo git NaCl 0,85%.
C Ly phn ln th hai ri ha tan phn vo git Lugol.
C B que g vo dung dch st trng.
y l knh ln 2 git phn.
C Kho st tiu bn di knh hin.

4. TIU CHUN CA MT TIU BN TT
Khng qu dy: phn nhiu s lm tiu bn c ti, che lp KST, kh pht hin.
Khng qu mng: t phn qu s khng tm thy KST, tr khi chng
qu nhiu.
Tiu bn c dy va phi khi thy c ch in trn t bo t di tiu bn.
Tiu bn khng c bt kh, dung dch phn khng trn ra quanh l knh.
5. KHO ST TIU BN DI KNH HIN VI
Kho st tiu bn phn bng vt knh x10, khi mun nhn r chi tit th chuyn sang vt knh x40.
Kho st mu phn theo hnh ch chi (zic zac) khng b st vi trng no.

Lu :
Nn nh sng va phi.
Mu phn c xt nghim cng sm cng tt, lu KST s cht hoc thay i hnh dng, kh xc
nh.
Mu phn tm trng giun, sn: khng qu 10 gi.
Mu phn tm n bo: khng qu 2 gi.
6. NHNG SAI LM NN TRNH
Pht phn khng u, ch dy, ch mng.
Nu pht phn long qu hoc c qu, nn b i, lm li pht phn khc.
y l knh lm tiu bn c bt kh.
Dung dch phn trn ra xung quanh l knh.
Qun khng t l knh ln pht phn th pht phn s chng kh, vt knh b bn v mu nhum s
nht rt nhanh.
Dng nc thng ha tan phn thay v dng dung dch NaCl 0,85%, nc thng s lm bin
dng hay hy hoi th hot ng ca n bo.
Dng nhiu nh sng qu. Nn t knh gn vi bn knh. Gim nh sng bng cch ng bt mng
chn sng hay dng knh lc mu xanh da tri ly nh sng.
7. CCH TR LI KT QU XT NGHIM PHN
Trn phiu tr li kt qu xt nghim phn, phi ghi cc ni dung sau:
c tnh ca phn: phn cng, mm, nho, c khun, lng,
Mu sc ca phn: vng, xanh, nu, en,.
Cc yu t bt thng thy c bng mt: cht nhy, mu, t sn,
K thut s dng: soi trc tip, k thut tp trung Willis,.
Kt qu:
+ m tnh: tm khng thy trng v bo nang ca KST ng rut.
+ Dng tnh, vit ra cc chi tit sau:
* Tn ting Vit v tn khoa hc ca KST.
* Trng, th hot ng, bo nang, u trng.
* Mt nhim trn tiu bn:

V d: Tm thy trng giun a (Ascaris lumbricoides): (+).
8. CCH X L DNG C DNG V BNH PHM
8.1. Bnh phm v que xt nghim
Sau khi xt nghim xong, cho l ng phn v que xt nghim vo dung dch st trng ri hp dit trng
trc khi b. Nu khng c l hp, c th nu si 30 pht hoc chn vo h su.
8.2. Dng c
Lam knh: cho vo dung dch st trng, hp kh trng, ra nc thng cho sch, ngm vo x bng,
ra sch, ngm trong dung dch acid sulfochromic 24 gi. Ra li bng nc thng, trng li bng nc ct,
sy kh.
L knh: ra tng t nh lam knh nhng ch sy kh 60
0
C.

CU HI LNG GI
1. M t k thut lm tiu bn phn soi trc tip tm KST.
2. Nu nhng sai lm c th gp khi lm tiu bn phn soi trc tip.
3. Trnh by cch ghi phiu tr li kt qu xt nghim phn.
K thut lm tiu bn phn

TT Thao tc Yu cu phi t
1
Chun b dng c v ha cht lm
tiu bn phn.
Dng c v ha cht y lm tiu
bn phn.
2 Ghi tn bnh nhn ln lam knh.
Ghi r tn hoc k hiu, i chiu vi tn
trn phiu xt nghim.
3
Nh ln lam knh 1 git NaCl 0,85% v
1 git Lugol.
Lng dung dch va , 2 git nc
khng qu gn hoc qu xa.
4
Cho phn vo git NaCl 0,85% v
khuy.
Lng phn va , khuy tan u, tiu
bn khng qu dy, khng qu mng.
5 Cho phn vo git Lugol v khuy.
Lng phn va , khuy tan u, tiu
bn khng qu dy, khng qu mng.
6 y l knh.
Khng c bt kh, bt nc. Dung dch
phn khng trn ra mp l knh.
7
t tiu bn ln bn knh hin vi. Tm
KST.
Soi ng theo quy trnh v quy nh.

Bi 5
K THUT TP TRUNG K SINH TRNG TRONG PHN

Tp trung KST trong phn l mt phng php thng quy chn on bnh nhim KST, cho php
pht hin thm mt s mm bnh m xt nghim trc tip khng pht hin c.
K thut tp trung KST trong phn c mc ch l tch KST ra khi cc cht cn b. C 2 cch: lm ni
KST v lm lng KST xung y ng nghim.
K thut ny c th s dng phn ti hoc c nh, phng i thp (100 v 400 ln) quan st tiu
bn, pht hin c trng v u trng giun, sn, bo nang n bo, trng nang ca trng bo t.
1. PHNG PHP LM NI
Trong phng php lm ni, dng mt dung dch hoc hn hp c t trng cao hn t trng ca cc bo
nang, trng giun, sn lm cho chng ni ln mt nc, cht b lng xung y ng nghim. Tiu bn soi
sch, t cn, nhng c mt s trng giun, sn khng ni m chm xung y nh trng giun a, trng c
np. V vy, vi phng php lm ni, cn soi lun cn khng b st KST.
1.1. K thut dng nc mui bo ha (Phng php Willis)
Phng php ny c dng tm trng cc loi giun, sn trong phn: trng giun mc (rt tt), giun
a, giun tc, trng sn di (dy) v sn di (dy) Hymenolepis sp.
Khng dng tm trng sn l, sn mng, u trng giun ln, bo nang v th hot ng ca n
bo.
a) Nguyn tc
Phn c ha tan trong nc mui bo ha. Trng giun, sn c t trng nh hn t trng ca nc
mui bo ha nn ni trn mt nc, dnh vo thy tinh (l knh) v c ly ra quan st di knh hin
vi.
b) Dng c
Knh hin vi
Lam knh
L knh
L penicilin hoc ng nghim 18 x 25mm
Que g
Hp Petri
Kp.
c) Ha cht
NaCl
Cn ethylic 95
0

Ether
Nc.
+ Dung dch nc mui bo ho:

Hoc cho mui vo trong nc cho n khi mui khng cn tan c na, ta c dung dch nc mui
bo ha.
+ Dung dch cn ether

* Ra l knh sch du, m bng cn ether :
C dung dch cn ether vo hp Petri.
C Cho l knh tng chic vo hp Petri, ngm trong 10 pht.
C Ly ra lau kh tng chic v ct trong hp Petri dng dn.
d) Quy trnh k thut
C Cho khong 5g phn vo l penicilin hoc ng nghim.
C vo l mt t nc mui bo ha, khong 1/3 l.
C Dng que khuy tan phn trong nc mui.
C Cho thm nc mui bo ha vo n khi mc nc ngang ming l.
C Vt b cc cn b ni ln mt nc.
Nh thm vi git nc mui bo ha vo l cho n khi mt nc cong vng ln (khng nc
mui trn ming l).
C y l knh ln ming l, trnh c bt kh gia l knh v mt nc.
yn trong khong 10 pht.
C Nhc thng l knh ln (l knh mang theo git nc mui mt di) v t ln lam knh.
Kho st tiu bn di knh hin vi.


Lu :
Nu thi gian ngn qu trng s cha ni ln.
Nu lu qu trng s ngm nc mui v chm xung y.
Thi gian di hay ngn ty theo cao ca ng nghim hoc chai l khi s dng. Tt nht nn th
thi gian trng ni vi ng nghim v chai l khc nhau.
1.2. K thut dng dung dch Sulfat km
Phng php ny dng tm trng cc loi giun v bo nang n bo trong phn.
Khng dng tm trng sn dy, sn l, sn mng, u trng giun ln. Khng dng vi phn c
nhiu m.
a) Nguyn tc
Phn c ha tan trong dung dch sulfat km bo ha, c t trng 1,18. Trng giun, sn c t trng nh
hn nn ni trn mt nc v c vt ra quan st di knh hin vi.
b) Dng c
Knh hin vi
ng nghim, ng ly tm
Gc
Khuyn cy vi trng
Que g
Lam knh
L knh
Kp.
c) Ha cht
Dung dch Sulfat km bo ho:

d) Quy trnh k thut
C Ha tan 5g phn vi 2 3ml nc trong ng nghim.
C Thm nc cho 10ml.
C Lc dung dch trn qua tm gc vo ng ly tm.
C Ly tm 2000 vng/pht trong 2 pht, b phn nc trong bn trn.
C Cho vo ng ly tm mt t dung dch Sulfat km, khuy u, tip tc cho thm dung dch Sulfat km
vo ng cho n cch ming ng nghim khong 2 3cm.
Ly tm 2000 vng/pht trong 2 pht.
C Dng khuyn cy trng ly phn ni trn mt dung dch ln lam knh.
y l knh.
2. PHNG PHP LNG
Tp trung KST trong phn theo cch lm lng cn, KST tp trung y ng nghim, c th pht hin
c nhiu loi KST, k c n bo, nhng bnh phm soi li cha nhiu cn hn.
2.1. Phng php lng trng lc
a) Nguyn tc
Mu phn c ha tan vi nc v lng t nhin. Trong cn lng c cha tt c KST c trong
phn.
Phng php ny cho php pht hin cc loi trng giun, u trng v trng sn mng (trong trng
hp nc c pha glycerin).
b) Dng c
Knh hin vi
Lam knh
L knh
Ly c chn
Que g.
c) Ha cht
Glycerin 0,5%.
d) Quy trnh k thut
C Dng nc hoc nc c pha glycerin 0,5%.
C Khuy u phn trong ly c chn.
C lng khong 1 gi.
C b phn nc ni bn trn.
C Thm nc, khuy u, lng 1 gi, gn b phn nc ni.
Lp li nhiu ln cho n khi nc bn trn trong hn.
C Gn b phn nc ni.
Khuy u phn cn, ly 1 hoc 2 git cn kho st di knh hin vi.
2.2. Phng php ly tm: K thut dng Formol Ether
a) Nguyn tc
Mu phn c x l vi formol bo qun KST c trong phn. Cht b c lc b, du, m trong
phn s c tch ra bi ether. Sau khi ly tm, cn lng xung y v cha tt c KST c trong phn.
b) Dng c
Knh hin vi
My ly tm
Lam knh
L knh
ng nghim
Gi ng ng nghim
ng nghim ly tm 15ml
Que g
Li lc kim loi
ng ht Pasteur
Cc c m.
c) Ha cht
Dung dch formol 10%.
Ether hoc ethyl acetat hoc xng.
d) Quy trnh k thut
C Dng que g ly khong 5g phn cho vo trong ng ly tm.
C Cho 7ml dung dch formol 10% vo ng ly tm.
C Ha tan phn v lc phn qua li lc vo ng ly tm khc hoc vo cc c m.
C Ly tm 2000 vng/pht trong 1 2 pht. Ht b phn nc ni. C th lp li nhiu ln cho n khi
phn nc ni trong.
C Cho 10ml dung dch formol 10% v 3ml ether vo trong ng ly tm.
y ng nghim bng nt cao su, trn u bng cch lc mnh trong vng 10 giy.
C M np cao su, cho ng nghim vo my ly tm, quay 2000 vng/pht trong 1 2 pht.
Ly ng nghim ra khi my ly tm, cht dch trong ng nghim c chia thnh 4 lp:
Lp trn cng: ether
Lp th 2: mt nt gm cc mnh vi cht bo dnh vo thnh ng
Lp th 3: formol
Lp th 4: cn cha KST.
Dng que g tch nh nhng lp cht bo ra khi thnh ng bng cch xoy theo hnh xon.
1 b 3 lp dung dch trn cng bng mt ng tc nhanh gn, dc ngc ng ly tm.
1 Dng ng ht Pasteur ly 1 git cn nh ln lam knh v kho st di knh hin vi (c th kho st
cn vi dung dch Lugol).
Lu :
Ether, ethyl acetat hoc xng l dung mi rt d bay hi v d chy nn xa ngun in, ngun la.
Bo qun cc cht ny trong bnh hoc chai l rng, ni thong mt. Khng t cc bnh cha Ether vo
t lnh, hi Ether s thot ra v gy n.

CU HI LNG GI
1. Mc ch ca phng php tp trung KST trong phn l g?
2. C bao nhiu phng php lm tp trung KST trong phn?
3. Nu u v nhc im ca k thut Willis v k thut dng formolether.
4. Anh (ch) s chn k thut no lm tp trung phn? Cho bit l do ca s chn la ny.
Xt nghim phn theo phng php tp trung Wills
Xt nghim phn theo phng php tp trung formolether

Stt Thao tc Yu cu phi t
1
tiu bn phn.
bn phn.
2 Cho phn vo l xt nghim. Lng phn ng quy nh 5 gram
3
nc mui bo ha vo l xt
nghim.
Khong 1/3 l.
4
Dng que g quy phn. Vt b cn to
(nu c).
Phn tan u trong nc mui bo ha,
khng c cn to.

thm mui bo ha vo l xt
nghim.
Nc y n ming l, hi vng ln
nhng khng trn ra ngoi.
5 y l knh ming l xt nghim.
Khng c bt kh, bt nc. Dung dch
phn tip xc vi l knh v tr cn i
trong thi gian 15 pht.
6
Nhc l knh ln v t tiu bn ln lam
knh.
Khng c bt kh, bt nc.
Soi knh hin vi tm k sinh trng. Soi ng theo quy trnh v quy nh.
1
tiu bn phn.
bn phn.
2 Cho phn vo l xt nghim. Ly nhiu ni trn khi phn.
3
dung dch formol 10% vo l xt
nghim.
Khong 7ml.
4
Dng que g quy phn. Lc dung dch
phn.
Quy cho phn tan u.
5
Quay ly tm 2000 vng/pht trong 1-2
pht, ht b nc ni.
Ht b nc ni khng lm xo trn cn.
Lp li giai on (5) nhiu ln. Phn nc ni phi trong.
6
dung dch formol 10% vo l xt
nghim. Cho thm ether.
Ly ng 10ml formol v 3ml ether.
7
y np ng nghim v trn u phn
vi formol v ether.
Np phi y kn v lc mnh ng nghim
trong 10 giy.
8
Quay ly tm 2000 vng/pht trong 1-2
pht. Ly ng nghim ra khi my ly
tm.
Ly cn thn, khng lm xo trn cc lp
dch trong ng nghim.
9 Tch b lp m.
Lp m c tch khi thnh ng cn
nguyn vn, khng b nt vn.
10
Trt b phn nc ni bng cch dc
nhanh ng nghim xung.
Sau khi b phn nc ni, cn khng
b xo trn.
11 Soi knh hin vi tm k sinh trng. Soi ng theo quy trnh v quy nh.

Bi 6
CC K THUT CHUYN BIT
PHT HIN K SINH TRNG NG RUT

Trong mt s trng hp, do c im sinh hc ring bit ca mt s loi giun, sn, n bo m cc
phng php xt nghim thng thng khng th hoc thnh thong mi pht hin mm bnh trong phn.
khc phc tnh trng ny, ngi ta nghin cu a ra cc phng php xt nghim c th ph
hp vi nhng c im sinh hc ca tng loi KST.
1. XT NGHIM TM TRNG GIUN KIM
1.1. Phng php GRAHAM (Phng php bng dnh trong)
a) Nguyn tc
Trng giun kim thng c tm thy cc np nhn hu mn, rt him khi tm thy trong phn. V
vy, pht hu mn bng keo dnh c dng thu thp trng giun kim. K thut ny phi c thc hin
bui sng sm trc khi tr lm v sinh hu mn.
b) Dng c
Knh hin vi
Lam knh
ng nghim
ng ht Pasteur
Bng keo trong
Cy li hoc mung di 10cm
Bng sch
Gng tay.
c) Quy trnh k thut
C Dng bng keo trong dn ln lam knh sao cho hai u lam knh u c ph mt on bng keo
khong 1cm.
C t ming lam knh dn bng keo ln cy li sao cho cnh nh ca tm lam knh cch b cy
li bng 1/3 chiu di ca tm lam knh.
C G bng dnh ra khi lam knh v cun vng qua u cy li, mt dnh ra ngoi.
C Gi cht tt c bng bn tay phi.
C Tay tri vch hu mn ca tr, tay phi cm cy li c dn bng keo n nh, lt qua lt li cy
li chung quanh ra hu mn.
Dn ming bng keo ln mt lam knh, dng bng kh v sch ch nh mit cht ming bng keo
xung mt lam knh.
C Kho st tiu bn di knh hin vi.
Ch : Phi lun mang gng tay trong sut qu trnh thao tc, trnh b nhim KST.

1.2. Phng php GRAHAM bin i
a) Dng c v ha cht
Knh hin vi
Lam knh
L knh
ng nghim
ng ht Pasteur
Que tm bng
Nc mui NaCl 0,85%
Gng tay.
b) Quy trnh k thut
C Dng que tm bng xoay chung quanh np nhn hu mn.
C Cho que tm bng vo trong ng nghim c cha 0,5ml NaCl 0,85%, ra k que tm bng vo trong
dung dch nc mui.
C Dng ng ht Pasteur ht nc mui ra lam knh, y l knh v kho st di knh hin vi.

2. TM U TRNG GIUN LN (PHNG PHP BAERMANN)
2.1. Nguyn tc
u trng giun ln c i tnh vi nhit v m cao v b thu ht v ni c nc m.

2.3. Quy trnh k thut
a) Lp t h thng dng c
C t phu thy tinh hoc plastic ln gi.
C t li kim loi ln phu.
C Lp ng cao su vo phu.
C Kp cht ng cao su li.
b) Thao tc
C Trn li kim loi t 2 ming gc.
C Nu phn lng, lt di ming gc 2 lp giy thm.
C khong 150g phn ti ln ming gc trn phu.
C vo phu nc m 45
o
C sao cho nc va sp ming phu v ngp phn.

C Dng bng n ri vo phu gi nhit ca nc.
yn t 1 3 gi.
C M kp kha ng cao su hng nc vo cc thy tinh.
Dng ng ht Pasteur ht nc vo ng ly tm.
Quay ly tm 1000 vng/pht trong 5 pht.
1 Gn b phn nc ni.
Dng ng ht Pasteur ht cn.
Kho st cn di knh hin vi tm u trng.
Mu phn c t 24 48 gi i khi thy u trng giun mc.
3. PHT HIN U SN DI (SN DY) TRONG PHN
Thuc iu tr sn di hin nay rt hiu qu nn vic tm u sn di trong phn cng khng cn thit
na. Tuy nhin, bnh phm phn cng nn c kim tra u sn v t sn mang trng nh danh loi.
Khi thc hin phi ly c b phn xt nghim.
Ha phn vi nc v cho qua h thng lc vi nhiu c mt li t to n nh (sp xp mt li t to
n nh).
Ly tm b phn nc ni, ly cn tm u sn bng knh lp hoc knh hin vi.

CU HI LNG GI
1. Cc phng php xt nghim phn trc tip c th pht hin c trng giun kim, u trng giun ln
khng? Ti sao?
2. M t thao tc thu thp trng giun kim.
3. K thut Baermann dng tm u trng giun ln, c th pht hin c u trng giun mc khng?
K thut Baerman

Bi 7
CY PHN

Cy phn c bit cn thit pht hin nhim nh giun mc, Strongyloides stercoralis v
Trichostrongylus spp. v nh danh k sinh trng.
u trng thng gp nht trong phn l giun ln.
1
tiu bn phn.
Dng c v ha cht y cy phn.
2 Lp t h thng cy phn. ng theo quy trnh.
3 2 lp gc ln li. Gc phi ph ton b mt li.
4 phn ln lp gc. Lng phn ln, nguyn b phn.
5 nc m ln phn. Nc m khong 45
o
C, va ngp phn.

6 yn t 1 - 3 gi. Lun gi nc m trong sut thi gian quy nh.
7
M kp kha ng cao su, hng nc
vo cc.
M kha nh nhng, khng nc vng
tung to ra xung quanh.
8
Dng ng ht Pasteur ht nc vo ng
ly tm.
Khng ri vi ra mi trng.
9 Ly tm. ng quy nh, nc khng vng ra xung quanh.
10
B nc ni. Nh 1 git cn ln lam
knh. y l knh.
Khng bt kh, khng trn nc ra quanh l knh.
11 t tiu bn ln bn knh hin vi. Tm KST. Soi ng theo quy trnh v quy nh.
Ty thi gian vn chuyn trong lng rut v iu kin ca bnh nhn, c th tm thy u trng c thc
qun phnh (rhabditiform) v him hn l u trng c thc qun hnh ng (filariform). Cng vy, nu v mt
l do no m phn b chm tng xut, trng phi v c u trng giun mc cng c th tm thy trong
phn.
C nhiu k thut cy: cy qua giy lc HaradaMori, cy giy lc hp thch, cy than.
1. K THUT HARADA MORI
1.1. Nguyn tc
Mt thanh giy thm c phn c cho vo ng nghim c t nc y. Nc thm dn ngc ln
thanh giy trong khi u trng giun di chuyn xung y ng nghim.

C Cho khong 3ml nc vo ng nghim.
C Tri phn ln ming giy thm, cch 2 u khong 2 3cm.
C t ming giy vo trong ng nghim sao cho nc chm vo u di ca ming giy nhng khng
ngp n phn.
C Ly giy bc u ng nghim li, chm l cho khng kh vo trong ng nghim.
C nhit phng th nghim, theo di hng ngy, t ngy 2 4.
Ht nc y ng nghim, nh 1 git ln lam knh.
C Nh thm 1 git dung dch Lugol, y l knh, 1 pht.
Quan st tiu bn di knh hin vi vi vt knh x10.
2. CY PHN TRONG HP PETRI
K thut ny ging nh k thut HaradaMori, ch khc l dng hp Petri thay v ng nghim.

C Ct giy thm thnh hnh ch nht 7 15cm.
C Cun cht giy thm quanh ming lam knh.
C Tri trn mt giy 1 gram phn.
C tt c vo y hp Petri c 10ml nc ct v trng v lun gi cho y hp c 1 lp nc trong
sut thi gian cy.
C nhit phng th nghim, theo di hng ngy, t ngy 2 4.
Ht nc y hp Petri, nh 1 git ln lam knh.
C y l knh v quan st tiu bn di knh hin vi.
3. KT QU
Nu kt qu cy dng tnh:
u gi th 48 c th thy:
+ u trng giun mc.
+ u trng giun ln.
T ngy th 2 n ngy th 4:
+ u trng giun mc.
+ u trng giun ln v giun trng thnh c, ci.

CU HI LNG GI
V nguyn tc, k thut cy phn HaradaMori c khc k thut Baermann khng?
K thut cy phn HaradaMori
1
Chun b dng c v ha cht lm tiu
bn phn.
Dng c v ha cht y cy
phn.
2 Cho nc ct vo ng nghim. Khong 3 - 5ml vo y ng nghim.
3
Tri phn ln thanh giy, cch u giy
khong 2 - 3cm.
Thnh mt lp u, khng dy qu,
khng mng qu.
4 t giy vo trong ng nghim.
Nc ch ngp u giy, khng chm
n phn.
5
Bc giy ming ng nghim, chm l
khng kh vo ng nghim.
Bc giy kn, chm nhng l nh rui,
kin khng chui vo c.

Bi 8
CC K THUT NHUM MU PHN

Phng php soi ti c gi tr rt ln trong chn on cc bnh KST ni chung, nhng khng cho
vic nhn dng n bo. V vy, cn phi b sung thm k thut nhum trong cc k thut chn on bnh
KST, c bit l i vi n bo. Nhum c nh l k thut quan trng nht xc nh nhim n bo
ng rut, v n gip cho vic phn bit cc b phn hnh th khc nhau nh vo s bt mu khc nhau ca
nhng b phn . Pht nhum ny c xem vi vt knh du ( phng i 1000 ln).
C nhiu cch nhum, ph bin nht l nhum Trichrome (Wheatley Gomori), nhum Haematoxylin
st v nhum Zeihl Neelsen ci tin.
1. NHUM TRICHOME (GOMORI WHEATLEY)
1.1. M t
Nhum Trichome l mt k thut nhum nhanh, n gin nhum n bo ng rut, nht l
xem cc trng roi, t bo ca ngi, nm ht men v cc KST gi trong khong 45 pht.
C th dng nhum mu phn ti hoc lu trong cn Polyvinyl hay phn mi c gn vi dung
dch Schaudinn.
Khng dng nhum mu phn c tp trung bng formol ether.
1.2. Vt liu, ha cht
ng cy nhit phng trong 2 - 4
ngy.
thi gian quy nh, khng nc
ngp phn, khng nc cn.
6
Tho b giy bc ming, ht 1 git nc
nh ln lam knh. y l knh.
B giy vo dung dch st trng.
Nc khng trn ra quanh l knh.
a) Thuc nhum Trichome
Thnh phn:

Cch pha v bo qun:
+ Cho 1,0ml acid acetic vo cc cht ni trn.
+ yn trong 15 30 pht nhit phng.
+ Cho 100ml nc ct vo, dung dch c mu tm.
+ Bo qun trong l thy tinh hoc nha nhit phng. Gi c trong 24 thng.
b) Dung dch cn ethylicIod
Thnh phn:

Bo qun trong l mu ti.
Khi dng pha long vi cn ethylic 70
o
cho ti khi c mu tr m.

Gi c khong 1 tun hoc ti khi no thy mu nht th b.
c) Dung dch Acid Cn ethylic

1.3. Quy trnh nhum
C nh tiu bn phn l bc rt quan trng trong qu trnh nhum c c tiu bn p. Trc y,
ngi ta thng c nh tiu bn phn bng dung dch Schaudinn v n nh hnh rt tt th hot ng v bo
nang ca n bo, nhng v dung dch Schaudinn c thy ngn, rt c, nn ngy nay ngi ta c th dng
dung dch Schaudinn c nh hay b ty ngi s dng.
a) Quy trnh c nh tiu bn bng dung dch Schaudinn
C Nhng ngay pht phn va lm xong vo trong dung dch Schaudinn, 30 pht hoc qua m, sau
ly tiu bn ra v nhng ln lt vo:

b) Quy trnh khng dng dung dch Schaudinn
Nhng tiu bn phn vo:

Quan st di knh hin vi vi vt knh du x100, nn s c mu hay xanh l cy, n bo c mu
xm nht hay l cy xanh nht.
1.4. Kt qu
Th hot ng v bo nang ca n bo c nhn thy d dng.
Trng giun, sn v u trng c th kh nhn din v thuc nhum m c.
C th nhn ra nm men, si t nm gi v cc bch cu.
Khng thy c trng bo t nh Cryptosporidium sp.
2. NHUM HAEMATOXYLIN ST (SPENCER MONROE METHOD)
2.1. M t
Phng php ny tt nhum n bo, t bo ca ngi, nm men, tinh th Charcotleyden, khng
dng nhum trng giun, sn hoc u trng v chng b nhum m en.
Phng php ny tt nhum tiu bn phn ti hoc phn c bo qun trong dung dch PVA
hoc SAF.
Phng php ny cng ging nh nhum Trichome. Quy trnh m t di y l c c nh bng dung
dch Schaudinn.
2.2. Vt liu
Knh hin vi
Lam knh
L knh
Que g
Bnh thy tinh dung dch nhum
Kp
Gng tay.
2.3. Ha cht
a) Dung dch A
Thnh phn:

Ha tan Haematoxylin tinh th vo 100ml cn ethylic 95
o
, yn ngoi nh sng trong 1 tun, sau
lc.
b) Dung dch B
Thnh phn:

Khi dng, trn ng th tch dung dch A v dung dch B, trc khi nhum t nht 3 4 gi.
c) Dung dch Cn ethylicIod
Thnh phn:
+ Iod tinh th 1 2g
+ Cn ethylic 70
o
100ml

Bo qun trong l mu ti.
Khi dng, pha long vi cn ethylic 70
o
cho ti khi c mu tr m.

Gi c khong 1 ti vi tun hoc ti khi no thy mu nht th b.
d) Dung dch Acid cn ethylic
+ Cn ethylic 90
o
99,5ml

+ Acid acetic bng 0,5ml
2.4. Quy trnh nhum


2.5. Kt qu
Th hot ng v bo nang ca n bo c nhn thy d dng.
Trng giun, sn v u trng c th kh c nhn din v thuc nhum m c.
C th nhn ra nm men, si t nm gi v cc bch cu.
Khng thy c trng bo t nh Cryptosporidium sp.
2.6. Cc vn thng gp
C nh tiu bn: nu khng c lm tt, c th kh nhn din n bo do b bin dng hoc khng
n mu.
Tiu bn lun c lm ro trc khi chuyn t bnh ny sang bnh khc trong qu trnh nhum bng
cch cm tiu bn theo chiu thng ng, cho 1 cnh lam knh chm vo giy thm rt cht lng.
3. K THUT NHUM ZIEHLNEELSEN CI TIN
3.1. M t
Cc trng bo t ng rut (Cryptosporidium, Cyclospora) kh c xc nh bng cch soi phn
trc tip. V vy, ngi ta dng phng php nhum to s tng phn gia mu ca KST v nn cn b
phn.
K thut nhum ZiehlNeelsen ci tin c dng da trn c im khng acid ca cc
KST ny. Nguyn tc ca k thut ny l nhum tiu bn phn bng carbon fuschin, sau
ty mu v nhum nn tiu bn bng mu xanh. Do c tnh khng acid nn cc trng bo t
gi li mu hng ca fuschin.
K thut nhum ZiehlNeelsen ci tin c th p dng cho phn ti, hoc phn c c nh trong
formol, hoc cc loi bnh phm khc nh dch ht t trng, mt, m (m).
3.2. Dng c
Knh hin vi
Lam knh
Gi ng lam knh
Que tm bng
Gng tay.
3.3. Ha cht
Methanol
Carbonfuchsin
Acid chlorhydric
Cn ethylic 95
o

Xanh Malachit 1% hoc Xanh Methylen 1%.
a) Pha Carbon Fuchsin
Dung dch A:

Dung dch B:

Trn 10ml dung dch A vi 90ml B li vi nhau.
Bo qun c 1 nm nhit phng.
b) Pha dung dch xanh Malachit

C th thay xanh Malachit bng xanh Methylen.
Bo qun c 1 nm nhit phng.
c) Pha dung dch cn acid chlorhydric

3.4. Quy trnh nhum

3.5. Cc vn thng gp
Nu lm pht phn qu dy, thuc nhum c th khng ngm vo tt c cc KST, c th lm sai lch
kt qu.
Khi ty mu, nu ty k qu (thi gian ty ko di hoc nng acid m) s lm cho KST khng cn
bt mu sau khi nhum.
Nu nhim nh, c th khng tm thy KST. Nn xt nghim 3 ln, mi ln cch nhau vi ngy
khng b st ca bnh.

CU HI LNG GI
1. Ti sao phi nhum phn?
2. C my phng php nhum phn?
3. M t phng php nhum Trichome.
4. Nu s khc bit gia phng php nhum Trichome v Haematoxylin.
K thut nhum ZiehlNeelsen ci tin
1
bn phn.
Dng c v ha cht y nhum
phn.
2 Lm pht phn mng trn lam knh. Pht phn mng, u.
3 kh ngoi khng kh. kh t nhin.
4
C nh bng methanol trong 5 pht.
kh ngoi khng kh.
ng thi gian quy nh.
5 Nhum vi carbonFuchsin trong 5 pht.
Ph thuc nhum pht phn, khng
nghing .
6 Ra lam knh vi nc.
Nhng vo su vo chu nc, trnh
cn bm vo tiu bn.
7
Ty mu bng cnacid cho ti khi mu
Nu ty k qu (thi gian ty ko di
hoc nng acid m) s lm cho KST

Bi 9
C LNG S GIUN BNG CCH M TRNG

Vic c lng s giun trong mt bnh nhn gip cho bc s lm sng bit cng nhim, quyt nh
kh nng ha tr liu v nh gi hiu qu thuc iu tr. Tuy nhin, ch c vi loi giun k sinh ngi mi
c s tng ng gia s lng trng trong phn v s giun c trong rut nh: Ascaris lumbricoides,
Trichuris trichiura, giun mc (Necator americanus v Ancylostoma duodenale).
Vic m trng ch l c on phi c thc hin mt cch cn thn.
C nhiu phng php m trng nh phng php Stoll, Brumpt, KatoKatz. Phng php c p
khng tri ra na. khng cn bt mu sau khi nhum.
8 Ra di vi nc chy
Dng nc chy va phi, khng qu
mnh.
9
Nhum vi 1% xanh Malachit trong 30 giy.
Ra tiu bn di vi nc. cho kh.
ng quy nh.
dng thng dng l pht dy KatoKatz.
1. K THUT LM PHT DY KATOKATZ
Pht dy KatoKatz c xem l mt k thut tt pht hin bnh sn mng Schistosoma mansoni v
mt s giun, sn khc rt hiu qu, hin ang c T chc Y t th gii chn lm phng php chun
pht hin trng giun, sn trong phn, c bit l giun a, giun mc, giun tc. Phng php ny khng dng
tm u trng, trng giun kim hoc n bo.
Pht dy KatoKatz l mt k thut c tnh cht nh tnh v nh lng do th tch phn c xc nh.
Lng phn dng ln nn d tm thy trng giun, sn hn xt nghim phn trc tip.
1.1. Nguyn tc: Phn c nh lng bng h ong c kch thc chun, tri ln lam knh v c lm
trong bi l knh bng giy cellophane thm glycerin.
1.2. Dng c
Knh hin vi
Que g
Li kim loi hoc plastic (60 105 mt li)
H ong phn bng kim loi khng g hoc bng plastic hay ba cng c mt th tch phn nht
nh
Que gt phn bng nha
Lam knh
Giy cellophane thm nc dy 40 50m, ct thnh tng ming c kch thc 22 x 35mm
Lc c np y bng
Kp
Giy v sinh hoc giy thm
Gng tay.
1.3. Ha cht
Xanh Malachit.
Glycerin.
Nc ct.
a) Pha ch dung dch xanh Malachit 1%

Bo qun trong l sm mu, nt kn.
b) Dung dch s dng
Thnh phn:

Trn u hn hp ni trn.
C th thay th xanh Malachit 1% bng xanh Methylen 3%.
Cc mnh giy cellophane c ngm vo dung dch ny 24 gi trc khi s dng.
C t mt mu phn nh trn giy bo (hnh 9.1a).
C n li ln mu phn sao cho phn lc qua li v t ln pha trn (hnh 9.1b).
C t tm h ong ln lam knh (hnh 9.1c).
C Dng que gt ly phn pha trn li cho phn y vo h ong, gt phn phn tha trn h.
C Nhc tm h ong ra sao cho phn trong h c gi li trn lam knh.
Ph ln phn mt ming giy cellophane c ngm dung dch glycerin mu, lau kh glycerin cn
trn mt giy cellophane (hnh 9.1d).
C Lt p tm lam knh ln mt mt phng cng, n xung nh nhng phn c tri mng u (hnh
9.1e).
Nhc cn thn tm lam knh bng cch trt nh nhng v mt bn, trnh lm rch hoc lm tch
mnh giy cellophane.
yn t 30 60 pht pht phn trong (nu mun lm trong tiu bn nhanh c th lam phn
vo t m 40
o
C hoc di nh sng mt tri trong vi pht).

1 Kho st tiu bn di knh hin vi.


1.5. c kt qu
Mu phn s trong, trng KST khng b trong s thy r trn nn mu xanh.
Phng php ny quan st d dng cc loi trng giun.
Lu :
Trng giun mc, trng sn mng (Schistosoma sp) v v mng nn d b lm trong v d v nu
lu. Trng mt v ch cn phi kh nhn dng. Do nn quan st sau khi ph giy cellophane 30 pht.
Phng php ny khng p dng cho phn lng.
K thut ny s dng phn ti, cn thn khi lm xt nghim.
1.6. m trng
Tnh s trng m c trong 1g phn:
Gi n l s trng trong X (mg) phn.
Gi N l s trng c trong 1g phn = 1000mg.
S trng m c trong 1g phn:

V d:
H ong c th tch: X = 50mg, n = 20.
S trng m c trong 1g phn l:

Trn thc t, kch thc h ong phn c tnh ton c th cha mt lng nht nh (X (mg)
phn):
+ ng knh 9mm dy 1mm cha 50mg phn.
+ ng knh 6mm dy 1,5mm cha 41,7mg phn.
+ ng knh 6,5mm dy 0,5mm cha 20mg phn.
Nh vy ch cn nhn s lng trng m c trong X (mg) phn vi h s (1000mg/X (mg)) s c
c s trng trong 1 gram phn.
1.7. nh gi
K thut ny n gin, nhanh, chnh xc, c th tin hnh hng lot trong iu tra v nghin cu. Tuy
nhin, vic c lng s giun k sinh ch c mt gi tr ht sc tng i v giun, sn khng theo nhp
u n, trng khng c phn b u trong phn. Phn c th c hay lng, lng phn thi ra hng ngy
c th nhiu hay t.
nh gi cng nhim ca giun a, giun tc, giun mc theo s lng giun k sinh v s trng
m c trong 1 gram phn theo quy c ca T chc Y t th gii (bng 9.1).
Bng 9.1
Loi giun
Nhim nh Nhim trung bnh Nhim nng
S lng
giun
S trng
/g phn
S lng
giun
S trng /g phn
S
lng
giun
S trng
/g phn
2. K THUT STOLL

C Cho dung dch NaOH N/10 vo bnh Stoll n vch 56ml.
C Ly khong 4g phn cho vo bnh cho n khi dung dch NaOH ln n vch 60ml.
C Cho bi thy tinh vo bnh.
C y nt bnh lc trn v u.
C qua m.
Ngy hm sau ly ra lc u trc khi ht mu.
C Dng ng ht ly 0,15ml dung dch phn nh ln lam knh y l knh, trnh c bt kh.
Tin hnh m trng trong ton b l knh.
2.3. Tnh kt qu
Gi n l s trng m c trong 0,15ml.
Gi N l s trng c trong 60ml (tng ng 4g phn).

S lng trng c trong 1g phn: n x 100.

Ty theo trng thi ca phn, nhn kt qu vi h s tng ng:
Giun a 20 < 5000 6 -24 5000 - 50000 > 25 > 50000
Giun tc < 1000 1000 - 10000 >10000
Ancylostoma
duodenale
20 < 3000 21 -100 3000 -10000 > 100 >10000
Necator
americanus
50 < 2000 51 -100 2000 - 7000 > 200 > 7000

Lng phn thi ra trung bnh mi ngy thay i ty theo tui:

T , tnh s lng trng thi ra trong mt ngy.
V d: Lng phn thi ra ngi ln l 200g:
S lng trng: n x 100 x 200
T s lng tnh ton c ta c th c lng s giun k sinh.
V d:
Giun a con ci mi ngy 200.000 trng.
S lng giun tnh ra t s lng m c:

Nu trong rut bnh nhn s lng giun ci v giun c bng nhau th s lng giun c trong rut bnh
nhn: X x 2.
Lu :
K thut m trng khng gip ta suy ra s lng giun, sn k sinh trong rut mt cch chnh xc v c
rt nhiu yu t lm sai lch kt qu nh:
Giun, sn khng trng theo mt nhp c nh: c ngy , c ngy khng.
S lng trng mi ngy cng khng ging nhau.
T l giun c/ci khng lun lun chnh xc.
Khng th nh gi ng trng thi ca phn, cch tnh h s ca mi tc gi cng khc nhau.
Lng phn thi ra mi ngy cng c nhiu thay i.

K sinh trng S trng /ngy T l c/ci Tng s KST
Giun a 200.000 1/1 2
Giun mc
(A.duodenale)
20.000 (50.000) 1/1 2
Giun mc
(N.americanus)
10.000 1/1 2
Giun tc 12.000 (50.000) 1/1,7 2,7
Sn l gan ln 1000 1
CU HI LNG GI
1. Anh (ch) cho bit mc ch ca m trng trong phn?
2. m trng c th tnh c cng nhim cho tt c cc loi giun, sn k sinh ngi? Ti sao?
3. So snh quy trnh m trng theo phng php Stoll v KatoKatz.
4. Theo anh (ch), hin nay c cn phi m trng trong iu tr bnh KST? Ti sao?
K thut KatoKatz

Bi 10
K THUT TM M TRONG PHN

1. MC CH
Th nghim tm m trong phn l nh gi hp th ca rut v tnh trng hot ng ca gan,
mt v ty.
S hin din ca cht bo trong phn, cn gi l tiu phn m, l biu hin ca ri lon tiu ha hoc
suy gim chc nng ca tuyn ty v mt.
Trng hp ri lon hp thu th s c nhiu acid bo v x phng, nu suy gim chc nng ca tuyn
ty v mt th c nhiu ht m trung tnh.
2. NGUN GC
Thc phm bnh nhn n vo:
T ng vt: m, tht ng vt, b.
1
bn phn.
Dng c v ha cht y lm
tiu bn phn.
2 Ly phn trn ry sng. ng quy nh
3 Cho phn vo h ong, t trn lam knh.
ng tc gn, sch, lng phn va
.
4 Nhc tm ong ra. ng tc gn, sch.
5
t giy Cellophane nhum mu ln
phn.
t ng v tr, ngay ngn.
6 Dng nt cao su p ln mt giy cellophan.
Phn phi dn u, mt giy tng i
phng.
7 tiu bn trong phng th nghim. thi gian quy nh.
T thc vt: du thc vt (du olive, du m).
3. CC DNG CHT BO
Acid bo, cht bo trung tnh, x phng (x bng).
4. HNH TH CHT BO DI KNH HIN VI
Acid bo: nhng ht trn nh cht bo trung tnh, hnh kim di, mnh hoc m v nh hnh.
Cht bo trung tnh: nhng ht trn, khng mu hay xanh l cy, vng, kch thc khng u.
X phng: hnh dng thay i, kh nhn dng, tng m ht, m v nh hnh, hnh kim ngn ng
ring l.
5. PHNG PHP BAILENGER
phn bit m trung tnh vi acid bo, ngi ta dng k thut nhum mu. C nhiu ha cht
nhum nhng ch nhng ha cht no gip phn bit c cc loi cht bo th mi c s dng v mu
phi bn, lu phai.
5.1. Nguyn tc
Da vo c tnh ca acid bo c i tnh vi Fuschine v acid bo, cht bo trung tnh n mu tm ca
Crystal violet.
5.2. Dng c
Knh hin vi
Lam knh
L knh
Que g
ng ht
n cn
Gng tay.
5.3. Ha cht

Lc trc khi dng. Bo qun trong chai c mu sm, nt tht kn.
C Dng que xt nghim ly mt t phn ln lam knh.
Nu phn c, c th pha long phn vi t nc.
C Cho 1 git phm nhum ln lam knh, trn u vi phn.
C y l knh ln, h nh qua ngn la.
5.5. c kt qu
M trung tnh c mu tm nht.
Acid bo c mu tm.
X phng c nhiu mu nhng phn ln c mu hng v cam.
Du paraffine khng bt mu.

CU HI LNG GI
1. Anh (ch) cho bit ngha ca m trong phn.
2. M t k thut Bailenger pht hin m trong phn.
3. M t cc loi cht bo khi soi ti v sau khi nhum.

Bi 11
K THUT TM MU TRONG PHN

1. MC CH
Tm mu trong phn chn on c xut huyt on no trn ng tiu ha. Xut huyt c th do
KST, do vim nhim, u bu, c th pht hin c bng mt, khi thy:
Phn c mu ti: c xut huyt on cui rut gi.
Phn en: xut huyt on trn ng tiu ha (rut non, d dy). Hng cu b tiu ha, khng cn
nguyn vn; huyt sc t c phng thch ra ngoi, ln vi phn (mu n). Trong trng hp ny, xc nh
xut huyt bng k thut tm huyt sc t trong phn.
2. K THUT TM HUYT SC T TRONG PHN BNG CHT
AMINOPYRINE
2.1. Nguyn tc
Khi huyt sc t tip xc vi hydrogen peroxide th oxygen c phng thch, oxygen ny s phn ng
vi cht Aminopyrine (Aminophenazone) lm hin mu.
2.2. Chun b bnh nhn cho xt nghim tm mu trong phn
Mt ngy trc khi lm xt nghim tm mu trong phn, bnh nhn khng c:
n tht.
Ung thuc c cha cht st.
nh rng mnh gy chy mu.
2.3. Dng c
My ly tm
ng ly tm
Que g
ng ong 20ml
ng nghim, gi ng nghim
Gng tay.
Lu : Dng c dng phi tht sch, khng c c vt mu.
2.4. Ha cht
Acid acetic 10%
Hydrogen peroxide 10% (pha trc khi dng)
Cn ethylic 95
o

Aminopyrine
Mu chng dng (dung dch 1% mu tan trong nc)
Mu chng m (nc ct).
C Pha dung dch aminopyrine (ch pha trc khi dng):
Cho 0,25g aminopyrine vo y ng nghim.
vo ng nghim 5ml cn ethylic 95
o
.

C Ly mt lng phn cho vo ng ly tm. Thm vo 7ml nc ct v nh tan phn.
C Ly tm 1000 vng/pht trong 5 pht.
C phn nc ni vo 1 ng nghim.
C Nh t t vo thnh ng nghim ng nc ni cc cht sau y:
10 git dung dch acid acetic.
5ml dung dch aminopyrine.
10 git hydrogen peroxide.
Ch : Khng trn ln cc cht vi nhau.
yn trong 1 pht.
C c kt qu trong vng 5 pht sau khi thm hygrogen peroxide.
2.6. c kt qu
Kt qu dng tnh th mu s xut hin gia 2 lp dung dch:
nht = phn ng dng tnh (+).
ti = phn ng dng tnh mnh (++).
m = phn ng dng tnh rt mnh (+++).
Khng c mu = phn ng m tnh ().

3. TH NGHIM PHT HIN MU TRONG PHN
L mt k thut pht hin mu trong phn nhanh, n gin, c nhy v chnh xc kh cao.
3.1. Nguyn tc
Huyt sc t trong phn gn vo khng th khng huyt sc t c dn ln thanh giy. Phc hp ny
c nhn thy nh cht nhum mu.
3.2. Dng c v ha cht
Thuc th mua trn th trng (FOB, ACON), gm c:
+ Thanh th trong bao gi
+ L nc pha long phn.
C Ly phn ca bnh nhn vo l kh, sch.
C Dng que xt nghim ly phn, t nht 3 ch khc nhau trn mu phn.
C Cho que phn vo l cha dung dch pha long phn v trn u.
C X bao ly thanh th ra.
C Nh 2 git (khong 90ml) dung dch phn ln thanh th vo ging c ch S.
c kt qu trong vng 5 10 pht.
3.4. Kt qu
Dng tnh: 2 vch
+ 1 vch chng
+1 vch ca mu th = kt qu dng tnh.
m tnh: 1 vch chng.
Nu ch c vch ca mu th hin ln = th nghim khng c gi tr.

CU HI LNG GI
1. Anh (ch) cho bit ngha s hin din ca mu trong phn.
2. M t k tm mu trong phn bng cht aminopyrine.

K THUT XT NGHIM TM K SINH TRNG TRONG MU
Xt nghim mu c th pht hin c cc n bo nh Plasmodium, Trypanosoma, Babesia,
Leishmania donovani v u trng giun ch.
K thut xt nghim c th soi ti hoc lm pht mu nhum.
Soi ti cho php thy c s chuyn ng ca KST trong mu.
Nhng vic nh danh thng phi nh n pht mu nhum c nh.
Pht mu c th lm t mu ton phn, ti, c hoc khng chng ng hoc cn lng t cc k thut
khc.

Bi 12
K THUT LM LN MU MNG
V GIT MU DY

K thut ln mu mng v git mu dy l xt nghim thng quy pht hin k sinh trng st rt
(KST SR) v mt s n bo k sinh trong mu khc. K thut ny cng c dng chn on bnh giun
ch h bch huyt.
Ln mu mng v git mu dy c nhng u im v nhc im, v vy trong chn on tm KST
SR thng lm c 2 loi tiu bn ln mu mng v git mu dy trn cng mt bnh nhn.
Ln mu mng:
u im:
+ Hng cu cn nguyn vn.
+ Hnh th KST p v in hnh, d nhn dng.
Nhc im:
+ Lng KST t do ch dng mt lng mu nh (vi ml)
+ Mt nhiu thi gian c lam mu v c th khng pht hin c.
Git mu dy:
u im: quan st 1 lng mu ln nn tp trung nhiu KST.
Nhc im: KST SR nm ln vi nhiu lp hng cu b ph v nn kh c nhn ra, i hi ngi
c lam mu phi c kinh nghim.
1. DNG C V HA CHT
Kim chch mu v trng
ng tim v trng
Bng thm nc
Gc sch
Lam knh kh v sch
Lam ko mu, c b tht phng
Hp ng tiu bn mu
Bt ch sp
n cn, khay men
Gng tay
Methanol c nh tiu bn
Cn Ethylic st trng 70
o
.

2. THI GIAN LY MU
Tm k sinh trng st rt:
Thi gian ly mu tt nht l trong thi gian ang ln cn st, v lc ln cn st KST SR tp trung
mu ngoi vi nhiu nn d thy.
Tm u trng giun ch bch huyt:
Ty thuc loi giun ch c chu k ngy hay m m ly mu. Vit Nam, u trng giun ch bch huyt
c chu k xut hin mu ngoi vi v m, nn thi gian ly mu tm u trng giun ch l vo khong t
22 gi n 3 gi sng.
3. CCH LY MU
Mu ton phn c ly t vt chch u ngn tay hoc t tnh mch.
3.1. Ly mu tnh mch
C Ly mu bng ng tim, bm vo ng nghim c sn cht chng ng EDTA 0,02g/10ml mu (hoc
mt cht chng ng khc). Nu tm giun ch th c th dng cht chng ng l Heparin (2mg/10ml mu)
hoc sodium citrate (0,05g/10ml mu).
C i vi mu ly t tnh mch vi cht chng ng nn lm tiu bn sm v nu qua 1 gi th hnh
dng ca KST SR c th b bin dng v mu d b bong ra v tri mt trong qu trnh nhum tiu bn.
3.2. Ly mu u ngn tay
a) V tr ly mu
Ngn tay th 3 hoc 4, bn tay tri.
u ngn tay, bn cnh ngn tay.
tr s sinh, c th ly mu gt chn.
b) Cch ly mu
C Phi dng phng php v trng, tt c dng c u phi c v trng trc.
C Ly mu t bn tay tri ca bnh nhn, nga lng bn tay ln trn, chn ngn tay th 3 hoc th 4.
C Dng mt ming bng v thm cn 70
o
lau sch u ngn tay. kh hay lau kh vi ming gc
kh trng.
C Dng kim chch mu v trng m vo ch chn vi mt ng tc nhanh v mnh 1 git
mu tro ra sau khi chch mu.
C Dng bng kh lau b git mu u v n c th ln vi cn cn st li.
Bp nh ngn tay lm chy git mu th 2.
C Cm 1 lam knh vo 2 cnh, chm nh ln git mu u ngn tay, mt git mu nh s dnh vo
ming knh gia lam knh lm ln mu mng.
Bp nh ngn tay nn thm mu, ly 3, 4 git mu vo 1 lam knh khc lm git mu dy (ln
gp ba git mu lm pht mu mng).
Dng bng thm cn lau sch phn mu cn li ngn tay.
Lu :
Khng nn chch mu khi st trng cha kh cn, mu chy lan ra.
Nu m kim nng qu, nn chch li, khng nn c nn cho mu chy ra.
Khi no khng ly mu na mi dng bng cn st trng ti ch ly mu.
C th lm git mu dy v ln mng mu trn cng 1 lam knh: v tr git mu dy khong 1/3 lam
knh v ln mu mng 2/3 lam knh.
4. KO LN MU MNG
C Cho 1 git mu ln 2/3 lam knh. t lam knh cha 1 git mu ln mt phng chc chn (hoc cm
trn tay).
C t 1 cnh lam ko mu ln lam knh c git mu thnh mt gc 45
o
.

C Ko t t cho lam ko mu chm vo git mu, cho mu lan theo giao tuyn ca hai lam knh.
C Ch cho mu lan ra gn ht cnh ca lam ko mu, y nhanh u v nh tay lam ko mu v pha
u kia ca lam knh cha mu.
C Nu git mu va th pht mu s khng ra n tn cng ca lam knh, m ngng li trc
khong 1cm, to ra ui mu.
kh t nhin, trnh bi, trnh cn trng n mu.
C Ghi tn bnh nhn phn dy ca ln mu mng, trc ch t git mu.
Ln mu mng t yu cu:
Ln mu phi mng u, khng c vt sc ngang, dc, khng loang l.
Ln mu c ui mng: Xem knh hin vi thy hng cu xp cnh nhau ch khng chng ln nhau v
cng khng cch xa nhau.
Nguyn nhn lm ln mu mng khng t yu cu:
Mu ly nhiu qu ko khng tt: tiu bn khng c ui mu.
Mu tri khng u: cnh lam ko mu khng phng hoc tip xc gia cnh lam ko mu v lam
knh ng mu khng kht.
Ko mu chm, ngp ngng hoc mu bt u ng: tiu bn s c nhng vt dy, sc.
Tiu bn c ch trng hoc l ch: lam knh bn, c m hoc rui, gin n.
5. LM GIT MU DY
C Ly git mu ln 1/3 cn li ca lam, 1 gc lam ko vo gia git mu. Xoay theo vng trn t
trung tm ra ngoi, theo mt chiu nht nh. Khi c c mt hnh trn ng knh t 1 1,2cm, xoay gc
lam ko i ngc vo trong ri nhc lam ko ln.
C kh lam mu trn mt phng, trnh bi v cn trng.
C Thng phi kh git mu dy trong khong 6 12 gi hay cch m, nhng khi cn gp c th
kh trong t 37
o
C, trong 1 gi.

Git mu dy t yu cu:
Phi u hoc mng dn v pha ba git mu.
Hnh dng tng i trn.
ng knh t 1 1,2cm.
Khng qu dy, git mu qu dy v qu to: lc kh mu s c nhng vt nt v d trc khi nhum.
Khng qu mng: git mu qu mng, nh (t mu), mt KST thp nn kh pht hin.
B dy thch hp, khi t tiu bn ln t bo lc cn t c th thy ch in.
C th lm git mu dy v ln mu mng trn cng mt lam knh hoc trn 2 lam knh khc nhau.
Khi lm tiu bn kp, ln mu mng v git mu dy trn cng 1 lam knh, hai git mu phi cch xa
nhau sao cho khi c nh ln mu mng bng cn th khng nh hng n git mu dy.


CU HI LNG GI
1. Anh (ch) cho bit gi tr ca git mu dy v ln mu mng?
2. Nu nhng u v khuyt im ca git mu dy v ln mu mng.
3. C th ly mu tnh mch lm tiu bn tm KST SR?
4. M t cch ly mu u ngn tay, cn ch khu no?
5. Nh th no l mt tiu bn mu mng p? Lm th no c tiu bn mu mng p?
6. Ti sao git mu dy bong ra khi nhum?
7. Nu nhng tiu chun ca git mu dy p.
Cch ly mu lm tiu bn tm k sinh trng st rt
1
Chun b dng c v ha cht ly
mu.
Dng c v ha cht y ly mu v
lm tiu bn.

K thut lm tiu bn ln mu mng
K thut lm tiu bn git mu dy
2
Ghi tn hoc s hiu ca bnh nhn ln
lam knh v i chiu vi phiu xt
nghim.
Tn bnh nhn c ghi r rng ln lam
knh v ng vi tn trn phiu xt nghim.
3 Chn v tr ly mu.
u ngn tay gia hoc p t hoc gt
chn.
4 St trng u ngn tay ly mu. Phi st trng ng quy cch v ng v tr.
5 Chch mu.
Phi ch da kh mi chch mu.
ng tc phi nhanh, dt khot, mnh va
phi 1 git mu tro ra da sau khi chch.
6 B git mu u. Sch, gn.
7
Bp nh ngn tay mu tro ra. Ly
mu t ln lam knh.
Ly 1 git, git mu trn, gn; lng mu
lm pht ln mu mng.
8
mu t ln lam knh.
Ly lng mu gp 3 ln lm pht git
mu dy.
1
Chun b dng c v ha cht ly
mu.
lm tiu bn.
2
Ghi tn hoc s hiu ca bnh nhn ln
nghim.
Tn bnh nhn c ghi r rng ln lam
3 Chn v tr ly mu.
u ngn tay gia hoc p t, hoc gt
chn.
5 Chch mu.
7
mu t ln lam knh
Lng mu ly va c ng knh
khong 3mm.
8
t cnh lam knh th hai tip xc vi
git mu theo gc 45
O
v ko mu.
Mu phi c tri u, ui git mu
nm cui lam knh, trn u.
9
git mu kh trn mt phng, trnh
bi, cn trng.
Git mu kh t nhin, c mu ng
nht, khng c bi, khng b nham nh.
1 Chun b dng c v ha cht ly mu.
lm tiu bn.
Ghi tn hoc s hiu ca bnh nhn ln Tn bnh nhn c ghi r rng ln lam

Bi 13
K THUT NHUM MU
TM K SINH TRNG ST RT

pht hin KST SR, tiu bn mu phi c nhum. Pht mu c nhum cng sm cng tt v
vic nhum cng gip ko di thi gian bo qun.
C nhiu cch nhum, trong c 2 cch nhum ph bin nht:
+ Nhum Wright, trong c sn cht c nh, vic nhum v c nh xy ra ng thi, do pht
mu dy phi c lm v hng cu trc khi nhum.
+ Nhum Giemsa, cht c nh v thuc nhum c tch ring, v vy, pht mu mng phi c
c nh vi Methanol tuyt i trc khi nhum.
Vit Nam, nhum Giemsa c dng nhiu nht.
1. DNG C
2
nghim.
3 Chn v tr ly mu.
u ngn tay gia hoc p t, hoc gt
chn.
5 Chch mu.
7
mu t ln lam knh.
Lng mu nhiu gp 3 ln pht ln mu
mng. Git mu dy khng qu gn pht
ln mu mng .
8
mt gc lam ko vo gia git mu.
Xoay theo vng trn t trung tm ra
ngoi, theo mt chiu nht nh.
Mu phi c tri u, git mu phi trn,
c ng knh t 1,2 -1,5cm.
9
git mu kh trn mt phng, trnh
bi, cn trng.
Git mu kh t nhin, c mu ng
nht, khng c bi, khng b nham nh.
ng ong c chia vi nhiu loi khc nhau: 10ml, 50ml, 100ml
ng ht nh git
Cc nh 50 250ml
Khay
Gi nhum tiu bn hoc bnh nhum
Gi ng lam knh hong kh tiu bn
ng h bo pht
Qut bn loi nh ( lm kh tiu bn)
Hp ng tiu bn
Kp.
2. HA CHT
Thuc nhum Giemsa (dung dch Giemsa m)
Methanol
Dung dch m phosphat pH 7 7,2.
Giemsa c cung cp trn th trng 2 dng: dng bt hoc dung dch pha sn.
2.1. Cch pha dung dch Giemsa m t dng bt
Thnh phn:

Cch pha:
Cho bt Giemsa vo trong ci vi mt t Glycerin, dng chy nghin bt Giemsa vi Glycerin cho tan
ht bt thuc nhum. Sau cho thm t t Glycerin vo nghin nhiu ln cho tan u, cui cng cho
Methanol vo.
Dung dch Giemsa m cn c ng trong chai thy tinh mu trung tnh, bo qun ch kh, mt v
khng c nh sng. Khi dng pha long dung dch Giemsa m theo nng quy nh.
2.2. Dung dch m
Nc trung tnh hoc hi kim (pH khong 7 7,2).
Dung dch m (dung dch phosphat, pH = 7,2) gm c:

Thi hn s dng dung dch m l vi tun. Khi no thy c cn th b.
3. K THUT NHUM TIU BN
3.1. Chun b tiu bn
a) Ln mu mng
Trc khi nhum phi c nh bng Methanol. Nhng lam mu vo bnh c Methanol hoc dng ng
ht nh Methanol ph kn ln mu mng, tiu bn kh.
b) Git mu dy
Trc khi nhum phi ph v hng cu, loi b huyt sc t bng dung dch nhc trng va phi
ph v hng cu nhng phi gi nguyn KST SR. Dung dch ty thng l dung dch Giemsa pha long
1%, dung dch m hay nc ct.
Ph dung dch nhc trng ln git mu dy, quan st ti khi mu hng ca mu tri i, li trn tiu
bn 1 git mu mu vng nht l c.

3.2. Quy trnh nhum tiu bn
a) Nhum thng quy
C t gi nhum trn khay nhum, khay ch phng, sau t lam mu ln gi nhum, mt c
mu ln trn.
C Pha dung dch nhum Giemsa 3%: 3ml Giemsa m + 97ml dung dch m.
C Ph kn dung dch nhum ln git mu.
C Thi gian nhum l 30 45 pht.
C Ra tiu bn bng nc ct hoc nc trung tnh: nhng su tiu bn nhum vo khay nc ra,
ly tiu bn ra nh nhng. Ra nh vy vi ln, n khi nc ra trong.
Cm tiu bn vo gi hong kh t nhin, mt mu quay xung di trnh bi.
C Ch khi no tiu bn tht kh mi soi di knh hin vi hoc ct bo qun trong hp ng tiu bn,
mun tiu bn kh nhanh dng qut, khng dng nhit .
Lu khi ra tiu bn:
Khng nn ht dung dch nhum i ri mi nhng tiu bn vo khay ra, nh vy cn thuc nhum
c th bm ln tiu bn.
Trnh ra tiu bn git mu dy di vi nc, v c th lm bong git mu.
b) Nhum nhanh
Quy trnh nhum ging nh trn, nhng pha dung dch nhum Giemsa 10%: 10ml Giemsa m + 90ml
dung dch m.
Thi gian nhum l 5 10 pht.
C th nhum tiu bn trong bnh nhum:
y dung dch nhum vo bnh.
Xp tiu bn mu c nh (git mu mng) hoc lm v hng cu (git mu dy) vo bnh
nhum.
4. NHN XT TIU BN NHUM TT
Tiu bn nhum tt, khi xem di knh hin vi thy nh sau:
Tiu bn sch, khng cn, khng bi.
Hng cu bt mu xanh tm hoc xanh da tri, hoc c mu hng nht. Nu c hng cu b nhim KST
SR, thy c nhng ht sc t v ht c hiu (ht Schuffner, ht Maurer).
Bch cu n nhn c mu xanh tm.
T bo cht ca bch cu lympho c mu xanh l nht.
Bch cu i toan c nhng ht mu ng r.
Bch cu a nhn trung tnh c nhng ht to nh, khng u, mu xanh l ti .
KST SR c hnh th r rng: nhn thng bt mu sm hoc ta, t bo cht bt mu xanh l, ht
sc t ca k sinh trng bt mu tm sm hoc mu nu en.
5. BO QUN TIU BN
Tiu bn nu lu li lu di cn bo qun tt. Khi soi KST SR xong phi lau sch du trn tiu bn bng
cch p tiu bn ln giy thm loi bt du, sau nh 1 2 git xylen ln pha trn git mu ri dng
khn vi mm, mng, sch, lau nh cho sch. Tiu bn ch khng c nh sng, tt nht l trong hp g.
CU HI LNG GI
1. Anh (ch) hy lit k nhng dng c cn nhum tiu bn mu bng Giemsa.
2. M t quy trnh nhum Giemsa tiu bn mu mng v dy.
3. Anh (ch) cho bit yu t no lm cho tiu bn nhum c p, thy r cc chi tit ca KST SR.
4. Nu tiu chun ca mt tiu bn nhum p.
Nhum tiu bn git mu mng
1 Chun b dng c v ha cht nhum
tiu bn mu.
Dng c v ha cht y nhum tiu
bn git mu dy.
2 C nh tiu bn mu bng cn
Methanol.
Nhng trn phn mu tri, khng lan ra
git mu dy.
3 Pha dung dch nhum. Pha ng cng thc.
4
Nhum tiu bn:
t tiu bn ln gi nhum hoc
vo bnh nhum.
Ph dung dch Giemsa pha

Tiu bn phi nm bng phng trn
gi, khng nghing.
Thuc nhum ph kn git mu,
Nhum tiu bn git mu dy

Bi 14
K THUT KHO ST TIU BN MU

1. CCH S DNG KNH HIN VI TM K SINH TRNG ST RT
long ln git mu. khng trn ra, chy xung khay.
5 Thi gian nhum. ng quy nh.
6 Ra tiu bn bng nc trung tnh,
hoc nc ct/ nc my.
Sch cn, khng c vng kim loi.
7 tiu bn kh trn gi . Git mu kh t nhin, khng b try st.
1
Chun b dng c v ha cht
nhum tiu bn mu.
Dng c v ha cht y nhum tiu
bn git mu dy.
2 Pha dung dch ty ph v hng cu. Pha ng cng thc.
3 Ty ph v hng cu.
Tiu bn mu sau khi b ph v hng cu
c mu trng hi c hoc c mu vng
nht.
4 Pha dung dch nhum. Pha ng cng thc.
5
Nhum tiu bn:
t tiu bn ln gi nhum hoc
vo bnh nhum.
Ph dung dch Giemsa pha
long ln git mu.

Tiu bn phi nm bng phng trn
gi, khng nghing.
Thuc nhum ph kn git mu,
khng trn ra, chy xung khay.
6 Thi gian nhum. ng quy nh.
7
Ra tiu bn bng nc trung tnh,
hoc nc ct/ nc my.
Sch cn, khng c vng kim loi.
8 tiu bn kh trn gi .
Git mu kh t nhin, khng b bong trc
hoc try st.
Tm KST SR cn phng i ln vi vt knh du (x100). Khi dng vt knh du soi lam mu, cn
ch :
nh sng.
Du soi phi trong.
Th knh, vt knh phi sch, khng b mc.
Trc y, c vi cp phi tt, khng b nhn.
Nng t quang ln st tiu bn.
Dng gng phng.
M rng mng chn sng.
2. C TIU BN MU NHUM
2.1. Cch c tiu bn mu nhum
Kho st git mu dy trc, ln mu mng sau.
Quan st tiu bn phng i nh (100 ln) trc qut tm giun ch. u trng giun ch him khi
c nhiu, thng ch c vi con trong mi pht mu.
Sau mi i sang phng i ln tm KST SR v n bo khc.
Trong mt tiu bn c th gp nhiu chng loi, v vy phi xem nhiu vi trng. i vi P.vivax,
thng thy nhiu giai on pht trin, cn P. falciparum, thng ch thy 1 2 giai on pht trin.
C th xem pht mu vi cc phng i khc nhau. Ty vo kh nng v kinh nghim ca ngi c
lam mu, vic kim tra pht mu thng mt 10 n 20 pht quan st 100 300 vi trng phng i
1000 ln.
2.2. c tiu bn git dy
Trong git mu dy, t bo mu tp trung nhiu nht gia. tm KST SR, tt nht nn c
phng i 1000 ln.
Kho st ton b git mu theo hnh ch chi xem c KST SR hay khng, phi quan st k vng
xung quanh git mu v nhng vng thng tp trung nhiu KST SR v sng, d xem hn.
Thi gian c thng t 5 n 10 pht (gn 100 vi trng vi vt knh du).
Hnh th KST SR trn tiu bn git mu dy v c bn ging nh hnh th KST SR trn tiu bn ln
mu mng. Tuy nhin do phng php lm tiu bn khc nhau, phng php nhum khc nhau nn hnh th
KST SR trn tiu bn git mu dy c khc hn mt cht, khng p bng tiu bn ln mu mng v KST
SR tp trung hn.
2.3. c tiu bn ln mu mng
Kho st phn ui v hai bn ra ca ln mu xc nh r loi KST SR.
KST SR thng hin din b hoc pha cui ca pht mu do qu trnh lm pht mu. ng thi,
ta nn kim tra KST SR phn cui pht mu, ni hng cu c tch ring ra. ni ny, hnh dng v kch
thc ca hng cu b nhim c thy r nht.
Khi xem phi kt hp 2 yu t: KST SR v hng cu b k sinh xc nh kt qu v loi KST SR.

3. MT S YU T GY NHM LN KHI KHO ST TIU BN ST RT
Trong tiu bn mu nhum Giemsa, c nhiu thnh phn trong mu, trong thuc nhum, nm, bi, c
th lm cho ngi c khng xc nh c. Nu nghi ng, nn lm li tiu bn khc hoc gi n nhng
ni chuyn su xc nh.
C th nhm cn thuc, mnh hng cu b v, nm, bi, tiu cu nh nm ring l vi th t dng
non.
Bch cu a nhn, m tiu cu nm t li d nhm vi th phn lit.
Hng cu nhn nhm vi th t dng gi, giao bo P.vivax.
Bch cu a nhn trung tnh thoi ha nhn d lm vi th phn lit, t dng gi P.vivax.
Bch cu n nhn, bch cu i toan v lympho bo khng n mu nhn d nhm vi th giao bo P.
vivax.
4. CCH TNH MT K SINH TRNG ST RT
Mt KST l s lng KST m c trong mt vi trng hoc mt th tch mu nht nh. Bit c
mt nhim KST l rt cn thit v gp phn tin lng bnh, nh gi hiu qu ca iu tr. C 2 phng
php dng tnh mt KST SR:
m s lng KST SR trong 1 microlit (1ml) mu.
H thng du (+).
4.1. m s KST SR trong 1l mu
T chc Y t th gii ly s bch cu = 8000/l mu lm chun.
m KST SR trong 1l mu tc l m KST SR trn 8000 bch cu (BC) trn git mu dy, sau
tnh ra s lng KST SR trong 1l mu.
k thut m c chnh xc, cn c cc iu kin sau:
+ Tiu bn nhum p.
+ Chn vng m: BC ri u, KST SR bt mu p.
+ Nn s dng 2 my m.
+ Nn m 2 3 ln ly s trung bnh.
Nu sau khi m c 200 BC, s KST SR m c = 10, th ngng m v tnh s lng KST
SR /200 BC.
Nu sau khi m c 200 BC, s KST SR m c = 10 th tip tc m KST SR cho 500 BC v
tnh s lng KST SR /500 BC.
Cng thc tnh:
S KST SR m c trn 200 BC = N.
BC/l mu = 8000

4.2. Tnh mt KST SR theo h thng du cng
nh gi mt nhim bng du cng (+) l mt cch m KST SR trn git dy n gin hn nhng
khng chnh xc bng cch m KST SR/l mu.
m KST SR trn tiu bn git mu dy:

5. CCH TR LI KT QU XT NGHIM TM K SINH TRNG ST RT
Nu tm thy KST SR th ghi kt qu theo th t nh sau:
Tn loi KST SR:
Cc th gp:
Mt nhim:

Nu tm khng thy KST th ghi:
Tm khng thy KST SR, hay ghi KST SR m tnh.

CU HI LNG GI
1. Anh (ch) ch n nhng yu t no khi s dng knh hin vi quan st tiu bn mu nhum?
2. Cho bit trnh t kho st tiu bn mu nhum.
3. Ti sao phi xem git mu dy trc ri mi xem git mu mng?
4. Anh (ch) cho bit nhng vt g c th gy nhm ln KST SR vi trn tiu bn mu nhum?
5. Theo anh (ch), vic m KST SR trong tiu bn mu c cn thit cho chn on bnh st rt? Ti sao?
6. C my cch m KST SR? Cch no thng c dng nht?
7. Anh (ch) ghi nhng ni dung no vo phiu tr li kt qu? Ti sao cn phi ghi y nhng ni dung
k trn?

Bi 15
K THUT XT NGHIM TM GIUN CH TRONG MU


1. CC K THUT DI Y C TH DNG TM U TRNG GIUN CH
Soi trc tip xem s di ng ca u trng. Bnh phm: mu, dch hch bch huyt hay cn lng nc
tiu.
Git mu dy, nhum Giemsa xem hnh th v cu trc nhn.
K thut Knott tp trung u trng.
Lc qua mng vi mng lc Nuclepore, tt trong nhim giun ch khi mt phi giun ch trong mu
thp.
Th nghim nhanh (test nhanh) chn on bnh giun ch: thao tc n gin v nhanh, c nhy
cao, nhng t tin v hin nay cha ph bin rng ri.
Sinh thit hch bch huyt tm giun trng thnh.
Th nghim huyt thanh hc.
nc ta, k thut thng c dng l git mu dy nhum Giemsa v k thut Knott.
Cch lm git mu dy nhum Giemsa: c bi lm git mu dy nhum Giemsa tm KST SR.
Di y l phn trnh by k thut Knott tm u trng giun ch trong mu.
2. K THUT KNOTT
K thut Knott l mt k thut tp trung giun ch trong mu, c dng trong nhng trng hp nhim
nh, c t KST. K thut ny s dng mt lng mu ln, hng cu b ly gii bi dung dch formol, sau
c ly tm. Giun ch s tp trung y ng nghim.
2.1. Dng c
Knh hin vi
Kim tim v trng
ng ng mu c Heparin
Lam knh
ng nghim hoc ng ly tm
My ly tm
ng ht Pasteur.
2.2. Ha cht
Dung dch formol 2%:

Trn k, lu gi trong l c np y. Thi hn s dng 24 thng.
2.3. Quy trnh thao tc
C Cho vo ng ly tm 10ml dung dch formol 2%.
C Thm vo 1ml mu ti hoc mu c cht chng ng.
C Trn k v ly tm 300 vng/pht trong 5 pht.
C B nc ni.
C Dng ng ht Pasteur trn u cn v ly 1 git nh ln lam knh.
y l knh ln git cn.
C Soi tm u trng giun ch di knh hin vi vi vt knh x10 v x40.
Nu tm thy u trng giun ch th lm tiu bn git dy, nhum Giemsa nh danh.
u im:
+ Thao tc n gin.
+ Cho kt qu nhanh.
+ R tin.
Nhc im: Khng thy c chuyn ng ca u trng, v n cht.

CU HI LNG GI
1. Anh (ch) cho bit nhng k thut dng pht hin giun ch?
2. M t quy trnh thao tc k thut Knott.

Bi 16
PHNG PHP CHN ON
BNG K THUT MIN DCH HC

1. I CNG
K sinh trng sau khi xm nhp vo c th bnh nhn c kh nng kch thch c th sn xut ra khng
th c hiu. V vy, cc nguyn l chung v min dch hc u c th p dng c i vi cc bnh k
sinh trng.
K thut min dch c s dng trong cc trng hp m phng php trc tip khng th lm c
nh:
+ Giai on mi nhim: KST cn non, cha trng (sn l gan, sn mng).
+ Trong giai on mn tnh, KST ng kn nh Toxoplasma.
+ Mt k sinh thp nh Trypanosoma.
+ KST nh v trong ni tng su nh bnh amp gan, go heo (ln) c, no.
+ Ng ct k sinh nh hi chng u trng di chuyn, KST dng u trng, khng bao gi tin n
giai on trng thnh v khng bao gi hon tt chu trnh pht trin nh Toxocara sp.
Trong nhiu trng hp, huyt thanh min dch hc t ra thc t hn, nht l trong cc trng hp m
cn ly bnh phm bng k thut xm ln.
2. K THUT MIN DCH HC
Phng php min dch hc p dng trong chn on bnh KST bao gm nhiu k thut nh kt ta,
in di, gn b th, ngng kt, min dch hunh quang, min dch phng x, min dch men.
Trc y, k thut min dch ch da vo khng nguyn pht hin v o lng khng th lun lu
trong mu v cc dch sinh hc. nhy v c hiu ca xt nghim ny ty thuc ch yu vo cht
lng ca khng nguyn, nu khng nguyn th th cho nhiu phn ng cho, dng gi, kt qu dng tnh
khng cho bit c bnh ang mc hay qua, v khng th gim rt chm, t vi thng n c nm. V
vy, vic bin lun kt qu gp nhiu kh khn v cn thn trng.
Vi nm gn y, chng ta c nhng b th nghim pht hin khng nguyn chn on mt s bnh
n bo nh bnh do Entamoeba histolytica, Giardia lamblia, Cryptosporidium parvum, Trichomonas
vaginalis v bnh giun ch Wuchereria, sn mng Schistosoma sp.
So vi nhm trn, cc k thut pht hin khng nguyn c gi tr chn on, kt qu dng tnh xc nh
bnh ang c.
3. K THUT MIN DCH MEN ELISA (ENZYME LINKED IMMUNOSORBENT
ASSAY)
K thut min dch men c nhiu loi: trc tip, gin tip, kp ch, cnh tranh. Hin nay, trong chn
on bnh KST, k thut ELISA gin tip c a dng v n c nhy cao, thao tc n gin, t tn ha
cht, khng i hi my mc t tin, c th lm hng lot. Phn m t di y l k thut min dch men
ELISA gin tip pht hin khng th trong mu bnh nhn.
4. K THUT MIN DCH MEN ELISA GIN TIP PHT HIN KHNG TH
Cc t vit tt:
KNKT = Phc hp khng nguyn khng th.
KKT = KT ca ng vt khng vi KT ca ngi.
Cng hp = KKT gn men.
4.1. Nguyn tc
Khng nguyn c gn ln ging nha, khi gp khng th tng ng s kt hp to thnh phc hp
KNKT. Phc hp KNKT c pht hin nh men gn vo KKT, men phn ng vi mt c cht v lm
i mu. Kt qu c th c c bng mt hoc my c ELISA.
4.2. Dng c v ha cht
ng nghim
Gi ng ng nghim
Micropipette c cc th tch khc nhau t 1l n 1000l
u nha gn vo Micropipette, 2 loi: mu vng (cho loi nh) v mu xanh dng (cho loi to 100
1000l)
Bnh nha ng nc ra (0,5 lt, 1 lt)
Giy thm p cho cc ging ro nc
My c ELISA (nu c th tt, khng c my th c kt qu bng mt)
B th nghim gm c:
+ Cc thanh nha gn khng nguyn
+ Dung dch ra
+ Dung dch pha long huyt thanh
+ Cng hp
+ Chng m
+ Chng dng
+ C cht
+ Cht ngng phn ng (acid hoc baz).
Thc hin theo hng dn ca nh sn xut.
Quy trnh gm cc bc sau y:
C Cho 100l huyt thanh bnh nhn, chng m, chng dng vo ging nha gn sn khng
nguyn. 37
o
C trong 1 gi.

C Ra 5 ln cc ging bng dung dch m PBS Tween 20.
C p cho ro nc.
C Cho 100l cng hp vo mi ging, 37
o
C trong 1 gi.

C Ra 5 ln cc ging bng dung dch m PBS Tween 20.
p cho ro nc.
C Cho 100ml c cht vo mi ging, ch cho mu xut hin (15 30 pht).
Cho acid hoc baz vo ngng phn ng.
c kt qu bng my c ELISA t ng hoc bng mt.
Kt qu dng tnh khi c hin mu, m tnh th khng c mu.

4.4. Bin lun kt qu
Ngng dng tnh c th khc nhau i vi tng loi bnh KST. Bin lun kt qu da trn ngng
dng tnh do nh sn xut a ra.
Cc k thut min dch pht hin khng th cho kt qu gin tip, khng c gi tr tuyt i.
Kt qu m tnh cng khng loi tr hon ton nhim KST, c th do mi b nhim, hoc b nhim qu
lu, hoc lng KST qu t c th kch thch c h min dch. Trong trng hp nghi ng trn lm sng
nn lm li th nghim. Xt nghim vi ln theo di bin ng ca khng th c gi tr hn l xt nghim
mt ln. Nu cn, phi kim tra li bng cc k thut khc.
Kt qu dng tnh cng khng khng nh hon ton b nhim KST v c th dng tnh gi do phn
ng cho gia cc loi KST.
V vy, cc phn ng min dch tm khng th khng th thay th hon ton xt nghim trc tip trong
chn on nguyn nhn bnh. Kt qu xt nghim, d m hay dng, cng cn c xem xt mt cch cn
thn, kt hp vi cc yu t dch t v triu chng lm sng.
5. TH NGHIM NHANH TM K SINH TRNG ST RT
Phng php ny c mc ch pht hin nhanh khng nguyn hay khng th chn on bnh st rt,
c thc hin di dng que nhng, khng i hi phng tin g khc ngoi b th nghim.
5.1. Th nghim Parasight F
Th nghim ny ch chn on bnh st rt do Plasmodium falciparum, bng cch dng khng th
n dng c hiu pht hin Protein giu Histidin (HRPII) do Plasmodium falciparum thi ra trong mu
bnh nhn.
a) Dng c v ha cht
ng mao qun
ng nghim
Gi ng nghim
B th nghim mua trn th trng:
+ Dung dch ly gii hng cu
+ Cng hp KKT Sulforhodamin B
+ Dung dch ra
+ Que nhng l thanh nitrocellulose.
b) Quy trnh k thut
Ly mu u ngn tay bng ng mao qun.
Cho 1 git mu vo trong 1 ng nghim c sn 3 git dung dch ly gii hng cu.
Nh 1 git mu b ly gii vo 1 ging th.
Nhng que th vo ging th, ch cho mu thm ln que th.
Nh 1 git cng hp vo ging nha.
Nh 2 git nc ra ln que nhng ra ht mu.
c kt qu:

Th nghim cho kt qu nhanh, trong vng 10 pht, c nhy, c hiu t 86 95% so vi lam
mu, nhng kt qu dng tnh khng xc nh c bnh ang pht trin hay lui.
5.2. Th nghim Optimal
C th pht hin c 4 loi KST SR gy bnh ngi.
Da vo vic pht hin men pLDH (Plasmodium lactate dehydrogenase), l mt loi men do k sinh
trng thi ra.
Men pLDH ch c sn xut ra bi KST SR cn sng nn th nghim ny c kh nng phn bit
KST SR sng hay cht.
a) Dng c v ha cht
ng mao qun
ng nghim
Gi ng nghim
B th nghim mua trn th trng.
b) Quy trnh k thut
Nh 1 git dung dch m vo ging th v 4 git vo ging ra trn phin nha.
Ly mu u ngn tay bng ng mao dn.
Nh git mu vo ging th.
Nhng que th vo ging th theo chiu thng ng, 10 pht cho mu thm ln que th.
Chuyn que th sang ging ra.
Sau khi ra, cc vch s xut hin r trn que th.
c kt qu:
+ C 1 vch : m tnh
+ C 2 vch : dng tnh vi 3 loi: P. vivax, P. ovale, P. malariae.
+ C 3 vch th dng tnh vi P. falciparum.
+ Khng c vch no: th nghim khng c gi tr.


CU HI LNG GI
1. Theo anh (ch), chn on bnh KST bng k thut min dch c th p dng cho nhng trng hp
no?
2. Ti sao k thut min dch men c s dng ph bin trong chn on bnh KST?
3. Nu nguyn l ca k thut ELISA.
4. M t k thut ELISA gin tip.
5. Nu nhng hn ch ca k thut Elisa pht hin khng th.
6. K thut min dch c thay th c phng php soi trc tip trong chn on bnh KST? Ti sao?
7. M t th nghim Parasight F.
8. So snh th nghim Parasight F vi pht mu trong chn on bnh st rt.
K thut ELISA
1
phn ng.
Dng c v ha cht y lm phn
ng.
2
Cho 100l huyt thanh bnh nhn,
chng m, chng dng vo ging
nha gn sn khng nguyn.
Ly ng khi lng, bnh phm tip xc
u vi mt ging, khng c bt kh.
3 37
o
C trong 1 gi.

ng thi gian theo quy nh.
4
Ra cc ging bng dung dch m
PBS-Tween 20.
Ra k nhiu ln theo quy nh.
5 p cho ro nc. Cc ging phi tht kh.

Bi 17
PHNG PHP IU TRA TIT TC

1. MC CH CA IU TRA TIT TC
Trong nghin cu v tit tc, cng tc iu tra tit tc l mt trong nhng cng tc c tm quan trng
c bit v nhng mc ch sau:
Bit c thnh phn loi v s lng c th ca mi loi tit tc.
Bit c s phn b ca tit tc theo a d.
6 Cho 100l cng hp vo mi ging. Ly ng khi lng.
7 37
o
trong 1 gi.

ng thi gian theo quy nh
8
Ra cc ging bng dung dch m
PBS - Tween 20.
c kt qu bng my c ELISA t ng
hoc bng mt.
9 p cho ro nc. Cc ging phi tht kh.
10
Cho 100l c cht vo mi ging, ch
cho mu xut hin.
Ly ng khi lng.
11 Cho acid hoc baz vo ngng phn ng.
Ly ng khi lng. Nh vo ging ng
theo thi gian quy nh.
12
c kt qu bng my c ELISA t
ng hoc bng mt.
c trong khong thi gian theo quy nh.
Bit c mt v bin ng s lng ca tit tc trong tng giai on i vi nhng iu kin thi
tit, kh hu khc nhau.
Tm hiu v sinh l v sinh thi ca tit tc.
Khm ph ra vai tr dch t ca tit tc.
T nhng hiu bit trn, tm ra nhng bin php phng chng v tiu dit nhng tit tc c hi vi
nhng loi thuc khc nhau v bit c liu lng thuc dit i vi cc loi tit tc.
Cn c vo kt qu iu tra trc v sau khi s dng thuc, bit c hiu qu ca cng tc dit tit
tc.
2. PHNG PHP BT MUI
2.1. Mui trng thnh
a) Dng c bt mui
ng nghim, ng ht, ng thng phong, mn (mng)
n pin
Bng khng thm nc
Bnh phun ha cht.
b) Ha cht
Ether hay chloroform
Pyrethrine
c) Phng php bt mui ban ngy
Bt mui bng ng nghim, ng ht, ng thng phong:
+ Mt tay cm ng nghim, chp thng ng vo ni mui u, lc nh mui bay vo pha trong
ng, ngn tay ci bt nhanh ming ng nghim, tay kia ly bng khng thm nc nt ming ng
nghim li.
+ Phng php ny ch bt c mui tng con v c th dng bt mui trong v ngoi nh, nhng
ch ti (bi cy, hc cy hay cc hc, cc x trong nh), dng n pin soi, nu thy mui, dng ng
thng phong chp mui.
Bt mui bng ha cht:
+ Thuc m (Ether hay Chloroform): phng php ny n gin, khi bt mui ch cn p ming ng
vo ch mui u, hi thuc xng ln lm mui ng ra v ri vo ng.
+ Pyrethrine dng kh dung: phun ha cht vo khng gian nh ng mui, c th bt tt c mui
trong nh. Phng php ny dng cho nhng nh kn o, t khe h.
Bt mui bng mn: dng mn lng bn ngoi mt khung g nh, mn ny c th trn cc lm cy,
bi c. Sau chui vo mn dng ng bt mui.
d) Phng php bt mui ban m
Phng php ny thng c kt qu tt, v a s cc loi mui hot ng v ban m.
Bt mui u trong nh: bng ng ht, ng nghim hay thuc m nh bt mui ban ngy.
Bt mui chung sc vt: dng n pin soi vo chung hay ngay trn sc vt, sau dng ng
nghim bt mui.
Bt mui bng mi ngi: mt hay hai ngi ngi ti mt a im, vn qun, tay o ln cho mui
t, khi mui u vo ngi dng n pin ri v dng ng thy tinh bt mui.
Bt mui bng by n: phng php ny li dng c tnh ca mui d kch thch v hp dn bi nh
n. n c treo ni bi cy c nhiu mui hay gn cc chung sc vt v n c treo sut t 4 gi
chiu n 6 gi sng hm sau.
e) Tnh mt mui
Mt mui trong nh c tnh bng cng thc:

2.2. Phng php bt b gy (cung qung)
a) Dng c
Vt li
Khay men
Mui mc /chn trng men
ng ht
Qu bp cao su
Chai l/ng nghim c 20 x 2cm.
b) Phng php bt b gy
Bt b gy bng vt li:
Phng php ny p dng cho cc thy vc rng nh sng, sui, ao, h. Dng vt li vt b gy:
t vt na di nc, na trn mt nc, ko mt ng di ri em vt ln, nhng vt vo khay men hay
chu nc trong th b gy ra. Sau dng ng ht, ht b gy b vo cc chai l.
Bt b gy bng mui hay chn trng men:
Phng php ny p dng cho nhng thy vc nh nh: h bom, nhng ni c nc ng. Dng mui
hay chn mc nc, sau dng ng ht, ht b gy cho vo cc chai l.
Bt b gy bng ng ht:
Dng ng ht di c qu bp cao su, phng php ny dng bt b gy trong cc hc cy, hc ni
qu nh khng th cho mui hay chn vo c. Ht tt c cc b gy v nc c trong hc ra khay trng
men, sau dng ng ht ht b gy ra chai l.
3. PHNG PHP BT B CHT
3.1. B cht trn chut
a) Dng c
By chut
Ti vi mu trng
Kp
X hay thau
Bn chi hay lc
ng ht
ng nghim hay l.
b) Ha cht
Ether hay chloroform
Cn ethylic 70
o
.

c) K thut bt b cht
C t by chut, chuyn chut vo ti vi mu trng v ct kn li b cht khi ri ra ngoi.
C Gy m chut bng ether hay chloroform, sau dng kp ln gp chut ra v t chut vo x
hay thau (c thnh cao, c vy nc t chung quanh thnh b cht dnh vo v khng b ra ngoi c).
C Dng bn chi hay lc chi cho b cht ri xung thau hay x.
C Dng ng ht nh hay que tre chut tht mnh ly b cht cho vo ng c cha cn ethylic 70
o
.

3.2. B cht trn ch, mo, sc vt nui
Dng bn chi hay ng ht bt b cht ngay trn mnh vt ch. Nu vt ch qu d, gy m vt ch
trc khi bt b cht.
4. PHNG PHP BT RP
C Tm bt qun o bn, c, c m hi, cc khe ging, t, bn, gh, khe sn nh.
C C th phun thuc dit cn trng xua rp ra khi ch n np.
C Tt c cc mu vt trn cho vo chai c cn ethylic 70
o
ghi r: ngy, thng, thnh phn loi, vt ch,
ni k sinh v a im bt.
5. PHNG PHP BT CH (CHY), RN
C Tm trn ngi tc, np qun o, lng.
C Dng lc dy chi tc, lng, hng ch, rn trn khay trng (hoc t giy trng).
C Dng kp gp ch rn cho vo ng nghim, y nt bng, dn nhn.
6. PHNG PHP BT RUI
Dng by rui bng nc ngt, qu thm.
7. PHNG PHP BT VE, MT
C Bt trn cc vt ch nh: ngi, ch, mo, g, chim,...
C Dng kp gp ve, mt cho vo ng nghim.
Ve thng b ln ngn cy c ch k ch i qua bm vo lng, qun o, V vy, c th dng mnh
vi trng lt trn mt cy c bt ve.
8. PHNG PHP BT M
Vt ch bt c cn kim tra v bt m ngay trnh m n no ri xung t.
chut: bt m tai, c quan sinh dc.
loi chim: bt m chung quanh m v lng ngc, di hai cnh.
loi b st: bt m di vy.
M bt c cho vo chai c cn ethylic 70
o
.

CU HI LNG GI
1. Anh (ch) cho bit mc ch ca iu tra tit tc.
2. C bao nhiu phng php bt mui?
3. Phng php bt mui no c dng ph bin nc ta?
4. M t phng php bt mui ban m.
5. M t cch bt b gy.
6. M t cch bt b cht, m, rp.

Bi 18
K THUT BO QUN
V CHUYN CH TIT TC

1. MC CH
Bo qun tit tc nh loi, lu tr, nghin cu v ging dy lu di. Cc mu vt bt c cn
c bo qun ng quy cch, trnh lm h hng hoc gy v mang v phng th nghim nh danh hoc
gi n cc phng th nghim chuyn su nh danh.
C rt nhiu phng php bo qun khc nhau ty theo loi, ty theo giai on pht trin ca cn
trng. Do , cn phi chn phng php bo qun cho thch hp vic nh loi c chnh xc v lu
gi lu bn.
2. DNG C
L nh th tch khong 30ml c np y
Chai c nt y
L c nt nha.
3. HA CHT
Cn ethylic 75
o
95
o

Formol 2 4%
Dung dch Barber
Dung dch Pampel
Dung dch Hall
Dung dch Mac Gregor.
3.1. Cn ethylic 75
o
95
o
: c s dng ph bin gi mu tit tc, c th thm 2 3% glycerin gi
cc loi cn trng nh: b gy, nhng ca mui, rui en, b cht, rui ct, ch, rn,
3.2. Dung dch Formol 2%: dng gi trng mui.
3.3. Dung dch Barber

Dung dch ny lm c th tit tc cng li, rt tt git cc cn trng nh v mt s loi u trng. Sau
khi ngm mu vt vo dung dch ny vi pht, vt mu vt sang cn v ngm trong vng 24 gi.
3.4. Dung dch Pampel

Dng bo qun u trng rui thuc h Muscidae.
3.5. Dung dch Hall: dng cho u trng rui.

3.6. Dung dch Mac Gregor

Dng gi b gy trnh c s cng v bo v cc mu sc.
5. K THUT BO QUN MUI
Cn phi tht cn thn, nh tay trong khi bt mui, trnh va chm nhiu lm mt hay hng cc c im
nh loi. Trc khi ct mui vo vt cha lu gi, cn ch cc im sau:
C Mui no ht mu, phi ch cho tiu ht mu ri mi chung vi cc mui khc.
C Mui c kh v ng nghim ng mui cng phi tht kh.
C Mi con mui c cho vo mt ng nghim ring.
C Mui bt tng nh phi hp ring, ghi r x, huyn, tnh v km theo phiu. Trong phiu phi
ghi r ngy, thng v a im bt mui.
6. K THUT BO QUN B GY V CC TIT TC KHC
6.1. B gy
C Dng nc nng 50
o
C 60
o
C git b gy, nhit ny b gy c th cht ngay v c th chng
s c gin ra.
C Sau chuyn chng qua chai c cc dung dch bo qun.
B gy bt c c th lu gi trong cc dung dch:
Cn ethylic 70
o
95
o
: thng dng nht.

Formol 2%.
Dung dch nc mui bo ha.
C Dn nhn trn vt cha b gy, trn nhn c ghi r ngy, thng, a im bt b gy.
6.2. Cc tit tc khc
Cc loi tit tc khc nh b cht, m, ch, rn, rp, mt,... c lu gi trong cc dung dch bo qun
nh:
Dung dch Pampel.
Cn ethylic 70
o
95
o
.

Formol 2%.
Km theo phiu ghi r ngy, thng v a im bt tit tc.
i vi m v b cht cn ghi thm chi tit bt t vt ch no.

CU HI LNG GI
1. Anh (ch) cho bit mc ch ca vic bo qun tit tc.
2. Trnh by cch bo qun mui.
3. Lit k cc dung dch bo qun tit tc v cho bit tc dng ca cc cht ny.
4. M t cch bo qun b gy.

Bi 19
PHNG PHP LM TIU BN TIT TC

Tit tc sau khi bt v c lm thnh tiu bn nghin cu v hnh th v nh loi.
C rt nhiu phng php khc nhau lm tiu bn tit tc t con non n con trng thnh. Quy trnh
gm cc bc:
Git cht mu vt.
Lm gin n mu vt nu cn.
Dng tiu bn.
1. CHUN B MU VT LM TIU BN
1.1. Dng c
L nh hay hp nh
Chu nh
K bng giy
Ct m.
1.2. Ha cht
C th dng mt trong nhng cht sau:
Chloroform/Ether
Carbon tetrachloride
Cyanid.
1.3. Mu vt: cc loi tit tc
Khi nhn mu vt, nu vt cn sng, ta phi git cht chng. Ty theo tng loi v tng giai on pht
trin ca tit tc m ta c cch git khc nhau. i vi nhng tit tc kh cng, ta phi lm mm v
dui chng ra.
1.4. K thut git tit tc
Nhng tit tc trng thnh thuc b Diptera v Hemiptera bt c ngoi a im, git ti ch ri
ng trong l nh hay hp nh.
i vi tit tc nui trong phng th nghim th cn phi c thi gian cho tit tc mnh m v cng cp
thng t 6 gi 24 gi, ri mi git bng nhng ha cht sau y:
Chloroform: thng dng nht.
Ether: cch dng ging chloroform.
Carbon tetrachlorid: dng thay chloroform v ethyl acetat, khng bt chy.
Dng ng nghim hoc l nh, cho giy thm vo, nh vi git chloroform vo ng nghim, cho tit tc
vo ng nghim, y kn ng nghim bng nt chai.
Cyanid: dng git cc loi tit tc ln nh gin. Sau khi tit tc cht th phi ly ra khi l cyarid
trnh s i mu ca c th.
1.5. Lm gin n cc mu tit tc
i vi nhng mu tit tc va git xong, ta c th lm tiu bn ngay.
i vi nhng mu b kh cng v do cc ni gi v th ta lm cho c th ca chng gin n ra
trc khi lm tiu bn chng khng b gy vn chn cnh khi dng tiu bn.
Cch lm gin n cn trng gm:
C Dng 1 chu nh ng ct m hoc ri nc t, pha trn c mt k bng giy, chu c th y
np kn. Ngoi ra, ta cn cho thm vo chu mt t cht thymol v phenol trnh nm mc.
C t mu vt ln k giy, y np. trong mt thi gian, lu hay mau ty thuc loi tit tc c kch
thc c th khc nhau, trung bnh t vi gi n 2 ngy.
C Sau tin hnh lm tiu bn nh mu vt ti.
2. K THUT LM TIU BN
2.1. Mu vt c kch thc nh: B gy
B gy v cc loi tit tc c kch thc nh khc c lm tiu bn gia lam knh v l knh. Trc khi
lm tiu bn, nu loi tit tc no c mu sc m, cn phi lm trong bng KOH 10%; cn vi loi tit tc
no c thn dy, cn phi p cho dp, ch p t t khng lm v mu vt. Di y l phn m t lm
tiu bn b gy.
a) Dng c
Que thy tinh
Hai kim cn trng
Lam knh
L knh
Nhn giy
B gy.
b) Ha cht
Dung dch dn lam knh:

c) Quy trnh k thut
C nh loi v tn b gy trc khi lm tiu bn.
C t b gy ln lam knh.
C Ly que thy tinh nh 1 git dung dch dn lam knh ln trn b gy.
C t l knh ln trn git dung dch dn lam knh.
C Nu cn, ly kim cn trng chnh sa t th thy r cc chi tit cn nh danh.
Nu c bt kh gia 2 lam knh th dng kim cn trng chnh cho ht bt kh.
C kh tiu bn hon thnh.
Dn nhn vo gc ca lam knh.
Ghi tn, ni bt, thi gian bt b gy ln nhn.
2.2. Mu vt c kch thc ln: Mui
Nhng tit tc c thn ln th khng th lm tiu bn gia 2 l knh, m c cm kim qua thn.
Sau khi cm kim, tit tc c gi trong ng thy tinh hoc ct trong hp kn. ng thy tinh phi tht
kh, sy k, y ng c cht chng mc, c ngn gi bng bng. Nt ca ng thy tinh phi tht kn v
gn bng Paraffine.
Nhng tit tc c thn hnh qu ln th c th ngm trong cn, khi cn ly ra quan st di knh lp
hoc knh hin vi. Sau khi nh danh tit tc ri, vit tn v gn nhn lm su tp.
a) Dng c
ng thy tinh nh c np
Kim cm: kim khng g, c th bng thy tinh
Bng
Bng phin
Nhn
Ming bc.
b) Ha cht
Ether
Paraffine.
c) Quy trnh lm tiu bn mui
Chun b dng c:
C Sy kh ro ng thy tinh nh c np.
C Cm kim thy tinh vo gia np.
Chun b mui:
C C th dng mui bt c hoc mui nui lm tiu bn, nhng nu dng mui c nui t b
gy cho n khi trng thnh th tiu bn s c gi tt hn.
C Nu bng mui cn mu th phi nui mt thi gian tiu ht mu ri mi lm tiu bn ( trnh b
mc).
C i vi mui cht v kh lu ngy:
+ mui ln mt tm giy thm ri t ln ming ng thy tinh c nc.
+ un ng thy tinh, hi nc bc ln ngm vo mui.
+ Mui gin n ri dng lm tiu bn (trnh gy, vn, kh).
Git cht mui:
C Bt mui cho vo ng thy tinh.
C t 1 ming bng tm ether hoc chloroform vo ming ng, (khng nn cho mui chm vo bng).
C th dng khi thuc l bng cch nh khi vo v y kn np li.
Lm tiu bn:
C mui cht ln tm bc.
C Sa cho mui nm nga, 2 cnh st vo bc, chn a ln.
C Cm np c kim cm m thng vo ngc gia (gia i chn gia). Khng nn cm qu su kim
l ln thn mui.
C Sau khi cm ta ly kim nh sa hai cnh v chn d quan st.
C Cm np c con mui vo ng (khng mui chm vo thnh ng trnh gy chn v cnh).
t ng mui vo t sy 56
o
C mui kh v dit trng.

C Sau 6 7 gi, ly ng ra ngui, m np ra, t nga np ln bn.
Cho nhn ghi tn, tui, a im, thi gian bt mui vo y ng.
Cho bng phin vo y ng, dy khong 1cm.
1 Nn cht bng thm nc sao cho mc ca bng ngang vi mc ca nhn.
un chy paraffine (khng si), nhng np ng thy tinh c mui vo paraffine, y ng thy tinh
li.

2.3. Tiu bn cc loi cn trng: ch, rn, b cht
C Git ch, rn hay b cht ri gi trong cn 90
o
(nu cn trng c mu th pha thm acid acetic 5%
trong cn 90
o
).

C Ngm trong dung dch KOH 10% khong 30 60 pht 50
o
C.

C Ra mu cn trng 2 ln vi nc sch.
C Chuyn sang cn 70
o
trong 30 pht.

C Chuyn sang cn 90
o
trong 10 pht.

Chuyn sang cn 100
o
trong 20 pht.

C Lm trong mu vt bng xylen hoc creozote trong 20 pht.
Chnh t th ca cn trng.
y l knh, trnh bt kh.
1 Dn tiu bn bng baume Canada.
kh nhit phng th nghim.
2.4. Tiu bn cc tit tc ln khc
K thut cm kim vo ngc tit tc c dng cho cc loi tit tc ln nh rui nh, rui tru, rui h
Calliphoridae, Oestridae. i vi vng nhit i nn chn kim khng g hoc kim bng thy tinh.
Cch cm kim:
C Dng kim m ng vo gia ngc tit tc v gi n cao 1cm cch u ca kim.
C Sau dng ming g ghim kim c tit tc, chnh cho cc b phn ca tit tc c ngang nhau.

CU HI LNG GI
1. Anh (ch) cho bit mc ch ca vic lm tiu bn tit tc v quy trnh lm tiu bn tit tc.
2. M t cch git mu vt tit tc ni chung.
3. Khi no cn lm gin n mu vt? M t cch lm gin n mui.
4. M t cch lm tiu bn b gy.
5. Trnh by cch lm tiu bn mui.
6. So snh cch lm tiu bn b cht vi tiu bn mui. Ti sao phi ngm b cht vo KOH?
7. M t cch lm tiu bn rui nh.


Bi 20
K THUT M MUI

1. MC CH
Mc ch ca m mui l nghin cu mt s tnh cht sinh l ca mui nh:
Xem mui c mang KST hay khng?
nh tui sinh l ca mui.
S ln ca mui.
Mui ht mu ngi hay mu sc vt.
2. DNG C
Mui
Knh hin vi
Knh lp
2 kim m mui
Lam knh, l knh
Bng gn thm nc
Khay men.
3. HA CHT
NaCl 0,85%
Cn ethylic 70
o

Ether (Chloroform).
4. QUY TRNH M MUI
4.1. Chun b mui
C Cn nh loi mui ngay trong ngy, nu bt mui vo bui sng th nh loi v m vo bui chiu.
Nu bt mui vo ban m th nh loi v m vo sng hm sau.
C ng nghim ng mui phi ch tng i m v ti.
C i vi mui bng cn mu phi ch cho tiu ht mu ri mi m.
C Gy m mui bng ether hoc chloroform, c th dng khi thuc l.
4.2. nh loi s b
Sau khi mui b m, t mui ln lam knh, cm kim vo ngc mui, nh ging bng mt thng hay
knh lp: Anopheles, Culex, Aedes, Mansonia.
4.3. nh h Sella
Khi mui ht mu no, tm ni u ngh v tiu ha mu. Mu c tiu ha n u th trng pht
trin n . nh gi qu trnh tiu ha mu bng cch nh h Sella.

4.3. K thut m mui
C Trn mt tm lam knh sch, nh 3 git NaCl 0,85%, git gia ln, hai git hai bn nh hn v mi
git cch nhau 1,5cm.
C Mui gy m, ct ht cnh v chn, t mui nm nghing vo git nc mui chnh gia, u
mui hng v pha tay phi, phn ui hng v pha tay tri, lng hng v pha trc, bng mui hng
v pha ngi m.
a) M tuyn nc bt
C t kim tri vo ngc mui, kim phi vo u mui, ko t t u mui ra khi ngc mui, lc
tuyn nc bt s ra theo u mui, l mt khi trong, chit quang.
C C th m ly tuyn nc bt bng cch ct u mui ra mt bn, sau dng kim nh ln ngc
mui gia t cng chn th nht v th hai, tuyn nc bt s li ra, dng kim ko tuyn nc bt sang git
nc mui bn phi, gt u mui ra ngoi.
b) M tch ly d dy v bung trng
C Xoay ngc tiu bn li, dng kim x rch mt t kitin t bng th 7 v th 8, t kim phi ln
bng mui, t kim tri ln t cui cng v ko t t sang tri, khi thy bung trng v d dy ra khi bng,
ly kim tch bung trng v ko sang git nc mui bn phi.
C Tch d dy ra khi rut, vt rut v tt c cc mnh kitin vn ra khi lam knh, ch li d dy
git nc mui chnh gia.
c) Kt qu
Xem tuyn nc bt c mang k sinh trng st rt hay u trng giun ch hay khng bng cch kho st
trc tip di knh hin vi, nu trong tuyn nc bt c thoa trng, thy c nhng hnh ging nh mng tay
ct ra xp li vi nhau thnh m v c chuyn ng hnh gn sng, mun xc nh chc chn, em pht ln
phin knh nhum Giemsa, ri em kho st di knh hin vi.
Tch d dy mui tm nang trng thnh d dy.
Tch trng t bung trng tnh tui sinh l ca mui. Quan st cui dy dn trng di knh hin
vi: l nhng t bo chit quang, c tng t dnh vo nhau. Mi ln mui li mt nt so to trn dy
dn trng (Nt Polovodova).

CU HI LNG GI
1. Anh (ch) cho bit mc ch ca vic m mui?
2. C nn m mui lc bng mui cng mu? Ti sao?
3. C th thy g tuyn nc bt, d dy mui?
4. Anh (ch) da vo yu t no tnh tui sinh l ca mui?
K thut m mui
1
Chun b dng c v ha cht m
mui.
Dng c v ha cht y m mui.
2 Gy m mui. Mui m su.
3 nh ging mui.
Nhn dng c mui thuc ging
Anopheles, Aedes, Culex hoc Mansonia.
4 nh h Sella. Xc nh ng giai on Sella.
5 M tch tuyn nc bt.
Tm c tuyn nc bt.
Tm c KST (nu c).
6 M tch bung trng. Tm c hai bung trng.

Bi 21
I CNG XT NGHIM VI NM

1. CHN ON BNH VI NM
Chn on bnh vi nm da vo triu chng lm sng v xt nghim tm nm ti phng th nghim. Cc
triu chng bnh gip cho nh hng chn on, cn xc nh bnh th cn phi da vo cc xt nghim
pht hin nm trong bnh phm.
Cc phng php kho st ca vi nm tng t cc phng php kho st ca Vi trng hc nhng i
hi nghim khc hn Vi trng hc. Sinh vin cn phi ht sc thn trng khi hc mn vi nm, v cc nm
sinh bo t, nhng bo t ny pht tn trong khng kh, ta rt d ht phi bo t nm hoc bo t nm bm
vo da. V vy, chng ta lun theo ng cc phng php v trng :
+ T bo v v bo v nhng ngi cng lm vic.
+ Gi c cc canh cy nguyn vn, khng b bi nhim bi cc tc nhn khc.
Bnh phm xt nghim tm vi nm: c nhiu loi bnh phm: lng, c, rn, v vy, cng c
nhiu cch ly bnh phm. Cn phi ly ng phng php bo m kt qu xt nghim c chnh xc.
7 Tm nt Polovodova. Xc nh ng tui sinh l.
8 M tm c ngc.
Tm c c ngc.
Tm c KST (giun ch nu c).
9 M tch d dy.
Tm c d dy.
Tm c KST (trng nang nu c).
10 Ghi phiu. Ghi ng quy nh.
Phng php pht hin vi nm gm c nhiu k thut khc nhau:
+ Kho st trc tip: bng mt thng v knh hin vi.
+ Nhum: tiu bn pht nm hoc sinh thit m (lt ct).
+ Cy nm.
+ Xt nghim sinh ha.
+ Phng php min dch v sinh hc phn t.
+ Tim vo th nui phng th nghim.
Kho st trc tip dng knh hin vi c th xc nh c chn on nu thy c nm trong bnh
phm, nhng khng thy nm th khng th loi tr nhim nm.
2. XY DNG MT PHNG VI NM
Mc ch ca phng xt nghim vi nm l chn on cc bnh do vi nm gip cho bnh nhn c
iu tr ng lc v kp thi.
Khi xy dng 1 phng vi nm, cn ch cc yu t sau y:
2.1. V tr
Phng xt nghim vi nm phi ni t c gi la, cch bit vi phng vi trng v siu vi trng. C th
xp chung vi phng k sinh trng.
2.2. Trang b
Phng xt nghim vi nm c th s dng tt c nhng trang b ca phng vi trng nh: knh hin vi, n
gas, n cn, n cc tm, khuyn cy, t , thuc nhum, v cn c thm:
Phng cy (ca la/gn my iu ha khng kh) hoc lng cy.
Kim cy nm.
Dao co, dao m, nhp, kp, ly mu th.
Dung dch KOH 10% 20%, xanh Lactophenol, mc tu.
n Wood quan st da, lng v tc.
Cc mi trng cy nm.
2.3. Bin php an ton trong phng nm
Ngi lm vic ti phng vi nm phi c nn (m), khu trang, o chong, gng tay,
Phi lau bn vi dung dch st trng trc v sau khi lm vic.
Ra tay bng x phng (x bng), dung dch st trng trc v sau khi kho st, cy bnh phm hoc
cy chuyn cc gc nm.
V sinh trong phng nm: khng c qut m ch lau vi dung dch st trng.
M n cc tm ngoi gi lm vic kh trng.
3. CCH LY V X L BNH PHM TM NM
Ly, vn chuyn v x l bnh phm rt quan trng trong vic xc nh nguyn nhn gy bnh. Ch
nhng bnh phm c ly v x l ng quy cch mi c th pht hin c tc nhn gy bnh r rng
nht. Bi v mt bnh phm c ly khng ng quy cch c th b nhim nhng chng nm thng tr v
gy kh khn trong vic bin lun kt qu do dng gi hay m gi.
3.1. Ly bnh phm
Bnh phm thng c ly trong phng m, ti ging hoc ni khc nn trch nhim ca phng
th nghim l c nhn vin n vi dng c thch hp. Tt c bnh phm u c xem l nguy him v phi
x l ht sc thn trng. L cha bnh phm khng c r r v ng trong mt hp nha chuyn n
phng th nghim. ng tim cha dch ht phi c rt kim ra v y kn trc khi vn chuyn.
Vi tt c cc bnh, bnh phm tt nht xc nh tc nhn gy bnh l ni nhim. Tuy nhin, nu
tc nhn gy bnh pht tn theo ng mu th c th ly mu chn on. Vic ly bnh phm cy nm
cng tng t nh cy vi khun. Nn hn ch dng tm bng ly bnh phm v nn ly bnh phm
su hn.
Nhng iu cn lu khi ly bnh phm tm vi nm:
Bnh nhn phi ngng iu tr trc khi ly bnh phm 7 10 ngy.
Khi ly cc loi bnh phm phi tun theo cc k thut v trng.
Phi ly bnh phm ng ch v nhiu c th lm cc k thut xt nghim khc nhau (soi ti
v/hoc cy).
3.2. Bnh phm
Mt s bnh phm do y bc s chuyn khoa ly.
Mt s bnh phm c ly ti phng xt nghim: da, tc, mng, m, mu.
Mt s bnh phm bnh nhn t ly: nc tiu, phn,
a) Tc
Chn nhng si tc pht hunh quang di nh sng n cc tm (n Wood).
Quan st u bnh nhn, chn nhng si tc mt bng, xp, mt mu, d gy hoc nhng ht mu en
bm tht cht vo si tc.
Nu c th c, cho bnh nhn bi thuc ln vng tc nghi nhim nm, qua m, hm sau gi
sch kh, chn nhng si tc cn gi mu thuc .
b) Da
Dng cn 70
o
st trng vng da b nhim nm, dng dao m v trng co da b vt thng cho cc
vy da rt ln tiu bn hoc vo hp Petri v trng.
Da rp ln nghi do lang ben: dng bng keo dn ln da, g ming bng keo ra, dn vy da ln tiu bn
hoc c th co da, ly vy quan st.
c) Mng
Dng cn 70
o
st trng mng nghi nhim nm, dng dao m v trng co mng hoc nhng cc li
xi ging nh bng ci di mng, hng bt mng ln tiu bn hay vo hp Petri v trng.
Nu mng c m: ly m bng que qun gn (bng) v trng.
d) m (m)
Ly m sng sm lc bnh nhn ng dy, sc ming tht sch, khc m vo l ming rng c np vn
v trng.
e) Pht hng
Lm pht hng bng que qun gn v trng v cho bnh phm vo ng nghim v trng.
f) M
Ly m bng que qun gn v trng ng tai, vim da, vim k, vim mng, cho vo ng nghim v
trng.
g) Mn bc
Dng kim chch v trng ly m.
h) Mnh gic mc
Do bc s chuyn khoa ly: co mt mnh vt lot hoc ghn cho vo l ng NaCl 0,85% v trng.
i) Mu
Cy tm nm: ly 5ml mu tnh mch, cho vo chai c 50ml BHI (mi trng tim, c hm) hay
Trypticase Soy, 37
o
C.

k) Cc dch: dch no ty, dch mng phi, nc ra ph qun,
Cc dch c ly v cho vo l cha v trng.
l) Mu sinh thit
Cc mu sinh thit c chia lm 2 phn:
Mt phn cho vo dung dch NaCl 0,85% lm xt nghim tm vi nm.
Mt phn cho vo dung dch c nh (Formol 10%, Bouin), lm tiu bn gii phu bnh l.
m) Phn
Ly mu phn vo l ming rng v trng.
n) Nc tiu
Tt nht l ly nc tiu gia dng, cho vo ng nghim v trng.
o) Huyt trng (dch m o)
Ly bng que qun gn v trng: pht ti cng Douglad hay cc mng trng, cho vo ng nghim c
dung dch NaCl 0,85% v trng.
3.3. Cch x l bnh phm
Bn cnh vic ly bnh phm chnh xc, vic vn chuyn v bo qun bnh phm ng quy cch cng l
yu t cn thit vi nm sng st. Trong cch x l bnh phm, ta cn ch n cc yu t sau y:
a) Thi gian. Bnh phm sau khi c ly ng cch, dn nhn v vn chuyn n phng th nghim nn
c x l cng sm cng tt.
b) Nhit
Vi nm c th b nh hng bi nhit nng v lnh, v vy, ch nn nhit phng trong vng 2
gi sau khi ly bnh phm. Hu ht bnh phm v bnh phm c kh nng nhim khun c th gi c
4
o
C; bnh phm l h thn kinh trung ng bo qun 30
o
C. i vi da, tc, mng, c th vn chuyn qua
ng bu in v nhng ni xa.
c) Khng nn x l nhng bnh phm nghi ng ly khng ng quy cch, nh l bnh phm ly v tr
khng ph hp, l cha b r r v kt qu xt nghim c th b sai lch, khng phn nh ng tnh trng bnh
l. Phi thng bo cho bc s lm sng ngay lp tc nu bnh phm b b, khng c x l; hoc thng bo
tnh trng nghi ng nu bnh phm tip tc c x l. Cc bnh phm b i u phi c tit khun trc
khi b.
d) Cn x l trc mt s bnh phm chn lc lm tng kh nng pht hin vi nm, quy trnh bao gm ly
tm ( tp trung vi nm), ly gii t bo mu ( phng thch vi nm khi t bo) v ct lt hoc ngm (
tng b mt tip xc ca bnh phm).
Ngoi vic soi trc tip di knh hin vi, pht hin khng nguyn v/ hoc lm phng php sinh hc
phn t, tt c cc bnh phm u c cy pht hin v nh danh vi nm.

CU HI LNG GI
1. Ti sao phi ly bnh phm ng quy cch?
2. c kt qu xt nghim c chnh xc, anh (ch) cn ch nhng iu g khi ly bnh phm
3. Lit k nhng bnh phm c th lm xt nghim tm nm?
4. M t cch ly bnh phm da, tc, mng.
5. C nn x l mu bnh phm ly khng ng quy cch? Ti sao?

Bi 22
K THUT XT NGHIM C BN TM NM

1. KHO ST TRC TIP
Khi quan st mu nm di knh hin vi, nu thy c nm trong tiu bn th c th xc nh ngi
bnh nhn b nhim nm. i khi ta c th nh danh c loi nm nu ta thy r cc bo t v nhng
cu trc c bit khc ca mt loi nm.
Nhng khi soi trc tip mt mu bnh phm m khng tm thy nm, ta cng khng th loi ngay kh
nng nhim nm. nhy ca mi phng php thay i ty theo loi bnh phm, v tr ly bnh phm, kinh
nghim ca ngi lm xt nghim.
1.1. Kho st vi NaCl 0,85%
NaCl 0,85% c dng vi bnh phm lng nh huyt trng, pht hng,
Quy trnh k thut:
C Pha long bnh phm trong dung dch NaCl 0,85%.
C Nh 1 git bnh phm pha long ln lam knh.
C y l knh v soi di knh hin vi vi vt knh x10 v x40.
C Kho st vi sng khc nhau (knh t quang cao, thp) tm vi nm dng si hay ht men.
1.2. Vi KOH 10% hoc 20%
Dung dch KOH hoc NaOH dng kho st cc bnh phm rn hoc m c (da, tc, mng, m,
da, mu sinh thit m,).
Dung dch KOH lm mm, cng gin cc t bo, tan cc ht du, m c trong bnh phm.
Quy trnh k thut:
C Trn tm lam knh sch, kh, nh 1 git KOH 10% hoc 20%.
C mu th vo git KOH, dm nh bnh phm nu l da hay trng tc.
C y l knh.
C yn 15 30 pht; hoc nu mun kho st ngay, h tm lam knh trn n cn (khng cho si).
C Kho st vi vt knh x10 v x40.
1.3. Kho st vi dung dch phm xanh Lacto Phenol Coton Blue (LPCB)
Thng p dng kho st cc la cy nm, Acid lactic gi vng cu trc vi nm, Phenol git nm,
xanh Coton nhum mu vi nm d quan st hn.
a) Quy trnh k thut
C t 1 mnh khun lc nm ln tm lam knh sch kh.
C Nh 1 2 git phm xanh LPCB.
C Dng kim cy ly la cy dm vo git dung dch phm xanh LPCB.
C y l knh.
C Kho st di knh hin vi.
Mun lu gi tiu bn, vin tiu bn bng keo sn mng tay.

b) Kt qu: Cc cu trc ca vi nm bt mu xanh tm ca Lacto Phenol Cotton blue.
2. PHNG PHP NHUM
2.1. Nhum mc tu
Thng dng kho st vi nm c nang, nht l vi nm Cryptococcus neoformans, mc tu khng
thm vo nang to mt khong trng, trng chung quanh t bo vi nm.
a) Quy trnh k thut
C Trn tm lam knh sch, kh, nh 1 git mc tu.
C Nh 1 git bnh phm, hay la cy ln git mc tu.
C t kim cy ngui, trn mc tu v bnh phm.
C y l knh ln git mc tu ha vi bnh phm.
Nu nn mc tu qu m c th lm long bt bng 1 git nc ct.
b) Kt qu
Nu trong bnh phm c Cryptococcus neoformans: t bo nm trn hay bu dc, bn ngoi c bao
bi 1 nang ln, nang khng thm mu, do sng ln trn nn en ca mc tu.
2.2. Nhum Gram
Thng dng kho st cc bnh phm dng lng, t nh: huyt trng, m, m, cn lng nc tiu,
pht hng,
a) Quy trnh k thut

b) Kt qu
Tt c vi nm bt mu Gram dng.
Tuy nhin c mt s vi nm khng gi mu tt, do cn dng phng php nhum Gram bin i.
2.3. Nhum Giemsa
Ch yu nhum bnh phm mu ngoi bin, ty xng vi nm trong t bo nh Histoplasma
capsulatum, k thut nhum ging nh nhum pht mu bnh thng.
2.4. Nhum khng acid
Nhum khng acid khi mun tm Norcardia.
a) Quy trnh k thut
C Pht bnh phm ln lam knh, c nh bng cch h nng.
C Ph Carbon fuchsin, trong 3 pht.
C b Carbon fuchsin.
C Nh Acid sulfuric ln tiu bn cho n khi khng cn mu ca Carbon fuchsin.
C Ph xanh Methylen, trong 1 pht.
Ra v hong kh.
C Kho st vi vt knh du.
b) Kt qu
Nocardia sp bt mu ca Carbon fuchsin.
2.5. Nhum PAS (Periodic acid Schiff)
Phng php ny thng c s dng cho cc mu m bnh phm tm vi nm dng si t v ht men.
a) Quy trnh k thut

b) Kt qu:
Vi nm bt mu tm.
Hin nay, ngi ta dng cht hunh quang pht hin nm nh thuc nhum Calcifluor trng pht
hin nm nhanh, k c Pneumocystis carinii, xanh Toluidine nhum m tm P. carinii.
Bng tm tt cc phng php nhum tm vi nm trong bnh phm
Phng php
nhum
Chn on Thi gian u im
Xanh Alcian Pht hin Cryptococcus neoformans 2 pht Tm thy trong DNT, c t c dng,
CU HI LNG GI
1. Phng php kho st trc tip c th p dng cho loi bnh phm no?
2. Anh (ch) cho bit cc loi bnh phm no thng phi dng KOH quan st v tc dng ca dung
dch KOH.
3. Trnh by quy trnh k thut lm tiu bn nm soi trc tip vi KOH.
4. Phng php nhum Giemsa dng cho loi bnh phm no v chn on loi nm no?
trong DNT. ngha l b vim mng no.
Acid nhanh Pht hin Mycobacteria v Nocardia
sp.
12 pht Pht hin Nocardia
v Blastomyces dermatitidis.
Kh nh
mu m.
Calcofluor Pht hin nm, P. carinii. 1 pht Pht hin nhanh nh pht
hunh quang.
i h
nhn d
Giemsa Nhum ty xng v pht mu
ngoi bin.
15 pht Pht hin
Histoplasma capsulatum.
Thng ch
Giemsa bin
i
Nhum m v nc ra
ph qun.
13 pht Pht hin th hot ng
ca Pneumocystis carinii.
Khng nhu
P. carinii
nm khc.
Gram Pht hin vi trng. 3 pht Thng dng pht hin vi
trng v vi nm.
Tt i v
nm khc khng
n mu nh
Nocardia sp
ging n
Mc tu Pht hin Cryptococcus neoformans
trong DNT.
1 pht Tm thy trong DNT, c
ngha l b vim mng no.
nh
KOH Lm trong nm. 5 pht
hoc hn
Pht hin nhanh nm. Ngi lm xt nghi
kinh nghi
Xanh Methylen Pht hin nm trong da. 2 pht Thng thm KOH
lm trong nm,
mu xanh gip pht
hin nm d hn.
M nn
nhn d
Methanamin
bc
Pht hin nm trong
mu m .
1 gi Tt pht hin nm. Cn ha ch
khng c s
k sinh trng.
Papanicolaou Pht hin t bo c tnh
trong dch tit.
30 pht C th pht hin c nm. Cn ha ch
c tiu b
PAS Pht hin nm ni chung. 20 - 25
pht
Nhum nm rt tt, thy r
si t nm v ht men.
Nocardia
B. dermatitidis
vt gi
Xanh Toluidin Nhum m v nc ra
ph qun. Pht hin nm,
P. carinii
25 pht Nhum bo nang P. carinii. Khng nhu
Khng th n
dng gn hunh
quang
Nhum m v nc ra
ph qun. Pht hin nm,
P. carinii.
45 pht Pht hin bo nang P. carinii. c hi
Wright Nhum ty xng v pht mu
ngoi bin.
7 pht Pht hin Histoplasma capsulatum. Thng ch
5. K thut soi vi mc tu c p dng cho cc loi vi nm no? M t quy trnh nhum v hnh nh ca
mm bnh trn tiu bn nhum.
6. K thut nhum PAS c p dng cho cc loi vi nm no? M t quy trnh nhum PAS v hnh nh
ca mm bnh trn tiu bn nhum.
7. Theo anh (ch)) nhum Gram c ch g trong chn on vi nm? Ti sao?
Xt nghim nm trc tip vi KOH
Phng php nhum mc tu

Bi 23
K THUT CY NM

1 Chun b dng c v ha cht lm tiu bn nm.Dng c v ha cht y lm tiu bn nm.
2 Nh dung dch KOH (10 - 20%) ln lam knh. Khi lng dung dch va theo quy nh.
3 t bnh phm vo git dung dch trn lam knh. bnh phm ngp trong dung dch.
4
Dm bnh phm trong dung dch. Bnh phm b ph thnh mnh nh nu l da ho
trng tc.
5
Lm trong bnh phm bng cch h tiu bn
trn n cn cho n khi dung dch bc hi.
Khng c si hoc cn kh.
6 Soi tiu bn tm nm. Thy c si t nm (nu c)
1
Chun b dng c v ha cht nhum tiu
bn.
Dng c v ha cht y nhum tiu bn nm.
2 Nh 1 git mc tu ln lam knh. Lam knh phi sch, kh.
3
Nh 1 git bnh phm, hay la cy ln git
mc tu, trn u.
bnh phm tan u trong dung dch.
4
y l knh ln git mc tu ha vi
bnh phm.
Bnh phm khng trn ra chung quanh. Khng c bt kh.
5 Soi tiu bn tm nm. Thy c nm ht men c bao dy (nu c).

1. MC CH
Kho st trc tip cho ta bit l c nm hoc khng c nm trong bnh phm, thng khng cho ta bit
chnh xc loi vi nm no gy bnh.
V d:
Nm ngoi da: ch bit c nm hay khng m thi.
Vim m o: soi ti huyt trng thy cc t bo ht men, ta ch bit l bnh nhn b nhim nm ht
men, ch khng bit tn loi nm ny. Nu thy t bo ht men v si t nm gi th ta c th bit tn l
ging Candida sp, ch khng xc nh c tn loi.
Mun bit chnh xc bnh do loi vi nm no phng xt nghim phi thc hin cy nm v nh danh.
Khi cy nm, ta phi chn mi trng cy nm thch hp vi mi loi bnh phm v vi loi nm nghi
ng.
2. MI TRNG DNG CY NM
C rt nhiu loi mi trng, bao gm nhng mi trng t ch v c sn trn th trng. Ty theo loi
bnh phm m ta chn mi trng thch hp:
Mi trng Sabouraud l mi trng c bn thng c s dng nhiu nht trong cy nm. Hu ht
cc nm u c th mc trong mi trng ny.
Mi trng Sabouraud + khng sinh thng c s dng cho bnh phm da, p xe v m di da.
Khng sinh ngn chn nhiu vi trng v lm gim tc mc ca nm lon nhim.
Khng sinh thng c dng: Chloramphenicol (<16g/ml); Gentamicin(5 100g/ml); Penicillin
(20U/ml); Streptomycin (40g/ml); Ciprofloxacin (5g/ml).
Mi trng tim c hm (BHI) b sung khng sinh: thng c s dng nht, dng cho bnh phm
ly t nhng ni khng v khun.
Thch bt bp c dng nh danh cc loi Candida v C. albicans.
Mi trng BHI v Sabhi thm khng sinh v 5 10% mu cu: Giu dinh dng, thng c s
dng pht hin nhng vi nm nh .
Mi trng sinh sc t (CHROMagar Candida, BD CHROMagar Candida v Candida ID) dng
pht hin v nh danh loi Candida.
Mt s mi trng nh thch Dextrose Sabouraud (SDA) thm Cycloheximide (0,5g/ml) c ch vi
nm hoi sinh nhng cng ng thi c ch mt s vi nm gy bnh nh Cryptococcus neoformans,
Trichosporon asahii, mt s loi Candida, Aspergillus v hu ht Zygomyces.
3. K THUT CY VI NM
3.1. Dng c
Knh hin vi
Kim cy trn hoc thng
ng nghim hoc hp Petri
ng ht Pasteur
Lam knh
L knh
T m.
3.2. K thut cy
Ty theo loi bnh phm v mi trng:
C Bnh phm t, cn lng cc dch, cy nm men:
S dng kim cy trn hoc thng ging kim cy ca vi trng.
Cy 3 chiu ging cy vi trng.
C Bnh phm kh nh: da, tc, mng., vi nm dng si:
Dng kim cy c bit cng.
Dm bnh phm hoc mng nm vo trong mi trng cy.
Mu tc, da: mi mu bnh phm, cy 3 5 ng mi trng.
C Cc bnh phm v trng: mu, dch no ty,.
Cy vo BHI hay thch mu trc khi cy vo Sabouraud.
4. TH NGHIM SINH NG MM
c dng phn bit Candida albicans hoc khng phi l Candida albicans.
4.1. Dng c v ha cht
Knh hin vi
ng nghim
ng ht Pasteur hoc khuyn cy vi trng
Lam knh
L knh
Huyt thanh ngi
Bnh phm hoc gc nm.
C Ly mt t nm t khun lc nm ha tan trong 1ml nc ct v trng to thnh mt huyn dch vi
nm.
C Nh 1 2 git la cy nm hoc la cy t mi trng Sabouraud lng vo mt ng nghim c cha
0,5ml n 1ml huyt thanh.
C 37
o
C t 2 4 gi.

C Dng ng ht Pasteur hoc khuyn cy ly mu cy nh ra lam knh.
C y l knh.
Soi tiu bn di knh hin vi vi vt knh x10 v x40.
4.3. Kt qu
Mc ra 1 chi di: Candida albicans.
Ny bp: khng phi Candida albicans.
Xt nghim ny c chnh xc cao. C th s dng huyt thanh ngi hay th c kh hay khng kh
b th u cho kt qu.
5. K THUT CY NM TRN KNH (S DNG CHO NM SI T)
5.1. Nguyn tc
Vi phng php cy nm trn knh: s lin h gia bo t, c cu sinh bo t, nhnh nm khng b xo
trn.
5.2. Mi trng
Thch Sabouraud hoc thch khoai ng: 15 20ml (cho 1 a thch).
5.3. Dng c v thuc th
Knh hin vi
Hp Petri, ng knh 90mm
Que thy tinh hnh ch U, V
Kim cy
Kp
Lam knh
L knh (tt nht c 18 x 18mm)
Nc ct v trng
Gc vi nm cn cy
Phm xanh Lacto phenol
Dung dch st trng.
C t lam knh ln que thy tinh un cong hnh ch U hay V.
C Tt c cho vo hp Petri, em hp v trng.
C thch Sabouraud vo mt Petri khc ngui. Dng dao v trng ct thch thnh khi 1 x 1mm,
t thch ln lam knh.
C Cy nm im gia mi cnh ca khi thch.
C H nh l knh ln ngn n y l knh ln khi thch.
Cho vo hp 1 3ml nc ct v trng.
C y np hp Petri, nhit phng th nghim.
Theo di la cy: khi thy nm mc, g l knh t ln tm lam knh c sn 1 git LPCB.
Dng kim cy ly khi thch trn lam cy b vo bnh nc st trng.
1 Lm thm mt tiu bn th 2: nh 1 git LPCB ln nm ni ming thch va c ly ra. y l knh
ln.
Lu gi tiu bn bng cch dn kn xung quanh l knh bng keo sn mng tay.
Ghi ch:
Nm hoi sinh mc: 3 7 ngy.
Nm gy bnh mc: 2 3 tun.
Khng nn cy trn knh cc loi nm nguy him.
V d: Histoplasma capsulatum

6. PHNG PHP DALMAU (S dng cho vi nm ht men)
6.1. Mi trng v dng c
Knh hin vi
Kim cy nm
L knh
Gc nm cn nh danh
ng nghim cha NaCl 0,85% v trng
Mi trng: thch bt bp.
C mi trng ra hp Petri.
C Ly gc nm cn cy ha tan vo ng nghim c cha NaCl 0,85% v trng (c th ly nm t la
cy lng).
C t kim cy ngui ly nm.
C Dng kim cy c nm vch su vo trong thch 2 ng song song cch nhau khong 1cm.
C t khuyn cy ngui, vch ng ch Z trn 2 ng cy.
H l knh trn ngn n st trng ri ph ln 2 ng cy.
C nhit phng 3 5 ngy.
Khi thy nm mc trn 2 ng cy, kho st trc tip nm trong hp cy di knh hin vi.
C th thch trn lam knh v cy ging nh cy trn hp.

7. NHIT V THI GIAN
Tt c mu cy c 30
o
C (1
o
C) bi v nhit ny thch hp cho hu ht vi nm gy bnh pht
trin. Nu khng th 30
o
C, nn nhit phng (gn 25
o
C). Trc y chng ta thng 4 tun,
nhng mt nghin cu gn y cho rng ch cn 3 tun l cho hu ht vi nm gy bnh pht trin, ngoi
tr da, tc, mng v bnh phm tm vi nm nh . i vi bnh phm tm nm men nh hu hng v m
o, ch cn 7 ngy. Nn kim tra mu cy t nht 2 hoc 3 ngy mi ln trong 2 tun u v hng tun
trong cc tun tip theo.
8. CCH QUAN ST MT CANH CY NM
8.1. Kho st la cy
Mt canh cy cn phi gi quan st trong mt thi gian t 1 3 tun. Trong thi gian ny, ta phi
thng xuyn theo di s tng trng ca nm mi ngy hoc 2 ngy 1 ln cho n khi c khun lc nm
hon chnh.
Tc mc ca khun lc nm:
+ Mc nhanh: t 2 5 ngy, thng l nm men hoc nm hoi sinh.
+ Mc chm: sau 1,5 n 2 thng mi c khun lc nm hon chnh: nhm vi nm gy bnh ngoi da.
V d: Trichophyton schoenleinii: mc chm.
+ Trung bnh: t 10 15 ngy.
+ Kt lun m tnh: sau 2 thng.
8.2. Kho st i th khun lc nm
a) Cu to mt trn ca khun lc
Cu to mt trn a dng:
Phng.
Nhn.
Nhn, gia cao ln nh ming ni la. V d: Trichophyton tonsurans.
Nhn nheo t trung tm ra ngoi vi. V d: Penicillium sp.
Ging nh rc bt: do vi nm sinh nhiu bo t. V d: Trichophyton mentagrophytes.
Ging nhung: do vi nm sinh si t nm ngn. V d: Microsporum canis.
Ging len: do vi nm sinh si t di. V d: Mucor sp., Rhizopus sp.
Ging sp. V d: Trichophyton schoenleinii.
b) Mu sc khm nm
Mt trn ca khun lc nm:
+ Mu sc rt a dng: trng vng, vng nu, xanh l cy, , en, ty theo loi vi nm.
+ Mt khun lc nm c th c 2 mu. V d Aspergillus sp.: Pha trong c mu xanh hoc nu, pha
ngoi c mu trng.
+ Trn mt khun lc nm c th ng nhng git nc. V d Penicillium sp.
Mt di ca khun lc nm:
+ Sinh sc t: c nhng loi vi nm khng sinh sc t. Mt s loi vi nm sinh sc t. V d:
Penicillium c mu vng v nu; Trichophyton sp. c mu , nu; Epidermophyton sp. c mu vng,
xanh l cy.
+ Phn b sc t:
* Chung quanh khun lc. V d: Epidermophyton.
* Lan ra ht ng nghim hoc a cy. V d: Trichoderma.
Ghi ch: kho st sc t nn so vi ng Sabouraud khng cy nm.
8.3. Kho st di knh hin vi
Dng kim cy ly mt mu khun lc nm. t mu nm trn mt lam knh c sn mt git LPCB.
Nu cn, ly hai kim ti mng ra, t l knh ln.
Quan st di knh hin vi vi vt knh x10, nu cn xoay qua vt knh x40.
Nu thy r bo t v nhng c cu c bit khc, ngi ta c th nh danh c canh cy nm.
Nhng nhiu khi ngi ta cn lm thm:
+ Cy nm trn ming knh xem v tr ca bo t.
+ Dng mi trng c bit cho sinh bo t hay sinh ra c cu c sc ca mt loi nm.
+ Kho st v tnh cht sinh ha hc hay c tnh khc ca nm.
+ i khi phi dng ti th phng th nghim.

CU HI LNG GI
1. Mi trng Sabouraud c thm khng sinh (Mycosel) c kt qu tt cho tt c cc loi bnh phm hay
khng? Gii thch.
2. Cy tm nm trong dch no ty dng k thut no?
3. K thut cy nm trn knh p dng cho loi bnh phm no? M t quy trnh cy nm trn knh
4. Anh (ch) cho bit mc ch ca k thut Dalmau v m t k thut ny.
5. Ti sao ta phi theo di mt canh cy nm thng xuyn v trong thi gian di?
K thut cy nm trn knh
K thut cy nm trn knh

1 Chun b dng c v ha cht cy nm. Dng c v ha cht y cy nm.
2
Cho que thy tinh un cong hnh ch U hay V vo hp Petri.
t lam knh ln que. y np hp Petri. Hp v trng.
Lam knh nm cn i trn que thy tinh.
3
Dng dao v trng ct thch thnh khi 1 x 1mm,
t thch ln lam knh.
Khi thch vung u, c t ngay
ngn gia lam knh.
4 Cy nm im gia mi cnh ca khi thch. Khng lm v thch.
5 H nh l knh ln ngn n y l knh ln khi thch. L knh c t ngay ngn, cn i ln kh
6
Cho vo hp 1 - 3ml nc ct v trng. y np
hp Petri - nhit phng th nghim.
Mc nc ph kn y hp Petri, khng ch
lam knh.
7 Theo di la cy. Theo di hng ngy.
8
Khi thy nm mc, g l knh t ln tm lam knh
c sn 1 git LPCB.
G v t nh l knh ln lam knh.
9
Gt b khi thch trn lam knh, cy vo bnh
nc st trng.
Khng nm vng ra xung quanh.
10 Nh ln ch nm mc 1 git LPCB. y l knh ln. Khng nc trn ra xung quanh.
11 Soi tiu bn tm nm. Thy c si t nm v bo t (nu c).
1 Chun b dng c v ha cht cy nm. Dng c v ha cht y cy nm.
2 mi trng ra hp Petri. Lp thch tri u, mt thch phng.
3 Ha tan nm vi NaCl 0,85% v trng. Nm tan u trong NaCl 0,85%.
4
Dng kim cy c nm vch su vo trong thch 2
ng.
ng vch sc nt, thng, cch nhau khong 1cm,
thch khng b v.
5
t khuyn cy ngui vch ng ch
Z trn 2 ng cy.
Thch khng b v.
6 H l knh trn ngn n, ph ln 2 ng cy. L knh c t ngay ngn trn cc ng vch.
7 nhit phng 3 - 5 ngy. Theo di nm mc hng ngy.
8
Khi thy nm mc trn 2 ng cy, kho
st trc tip nm trong hp cy di knh hin vi.
Thy c nm.

Bi 24
HNH TH U TRNG V GIUN TRNG THNH

1. GIUN TRNG THNH
1.1. Cn c nh loi
nh loi giun, sn trng thnh da vo cc c im:
Hnh dng
Kch thc
Cu to.
1.2. c im ca lp giun
Giun c hnh ng, trn, di t vi milimt n vi chc centimt, i xng hai bn.
Thng c mu trng c, c mt ng tiu ha trn vn.
Thn giun c bc bn ngoi bi lp v hyaline.
Di lp v l biu m (epithelium) v mt lp c dc. Lp biu m nh vo pha trong to thnh 4
g dc c th: 1 g lng, 1 g bng v 2 g bn. G lng v bng mang dy thn kinh, 2 g bn mang ng
bi tit.
phn u c ming vi cc b phn nh mc, rng, bn dao, gai.
Xoang ming c dng ng v c th bin i thnh b phn ht ty theo tng loi.
Thc qun hnh ng, i khi phnh phn cui, l c im nh danh trong vi trng hp.
Rut l ng dp ni thng t thc qun n hu mn.
H bi tit gm 2 ng chy dc theo hai bn thn v 1 ng ni ngang pha gn u, ra ngoi bng
l bi tit khong thc qun.
H thn kinh gm 1 vng ni cc hch thn kinh quanh thc qun. T vng ny, 6 si thn kinh i
pha trc v 6 si thn kinh chy v pha sau.
C 2 gii ring bit. Con c lun nh hn con ci, ui cun li hay xo ra nh ci ti hnh chung.
Con ci c ui thng.
C quan sinh dc c nm 1/3 sau ca c th, l ng n hnh chp hoc ng xon cun gm tinh
hon, ng dn tinh, ti cha tinh, ng phng tinh, tn cng bng c quan giao hp. C quan giao hp c th
l 1 hoc 2 gai giao hp. vi loi, c quan giao hp c dng cnh hoc dng ti.
C quan sinh dc ci c th l cu trc n hoc i gm bung trng, ng dn trng, ti nhn tinh, t
cung, m o v m mn m ra ngoi bng l sinh dc na thn trn, pha bng.

1.3. Giun a (Ascaris lumbricoides)
Thn hnh ng, u n, thon 2 u, c v bc ngoi dy, cng, c vn ngang.
C mu trng hoc hi hng hoc nu nht.
u giun thun nh, c 3 mi xp cn i (1 mi lng v 2 mi bng).
Giun c di 15 17cm, ng knh 0,3 0,4cm, ui cong, l hu mn mt bng, gn cui thn,
c 2 gai giao hp.
Giun ci di 20 25cm, ng knh trung bnh 0,5 0,6cm, ui thng, hnh nn. L sinh dc nm
khong 1/3 trc ca thn mt bng.

1.4. Giun tc (Trichuris trichiura)
Thn chia lm 2 phn: Phn u mnh nh si tc v chim gn 2/3 thn, phn cn li hnh ng, to,
cha cc c quan sinh dc.
Giun tc c mu hng nht.
Con c di 30 40mm, ui cong, cui ui c 1 gai sinh dc nm trong bao gai.
Con ci di 30 50mm, ui thng v bu, l sinh dc nm 2/3 thn k t u, c quan sinh dc
gm c 2 bung trng hnh ng cun li nh l xo v t cung.


1.5. Giun mc (Ancylostoma duodenale, Necator americanus)
Giun mu trng hoc hng.
Mu xm, nh nh si ch.
Kch thc: 7 x 10mm x 0,4 0,5mm.
u giun mc c bao ming, thc qun hnh ng:
+ Ming c 2 i rng u: Ancylostoma duodenale
+ Ming c i dao ct: Necator americanus.
Con c di 8 11mm, ng knh thn di 0,5mm, u cong, ui xo ra thnh ti, c nng bi
nhng ng gn, c 2 gai giao hp di v mnh.
Con ci di 10 13mm, ng knh thn 0,6mm, u cong, ui thng hnh chp.


1.6. Giun ln (Strongyloides stercoralis)
Hu ht giun ln k sinh l giun ci:
Kch thc 2mm x 50mm.
ng knh khng thay i, nh nh si ch trng, gn nh trong sut.
ng thc qun hnh ng.
ui nhn.
m h cch u mt khong di hn 1/3 thn.
Trng sp thnh tng dy trong t cung.


1.7. Giun kim (Enterobius vermicularis)
Mu trng.
Di khong 1cm.
Ming gm 3 mi, khng c bao ming.
V bc ngoi dy ln phn u v dc 2 bn thn.
Phn cui thc qun n thnh hnh bu trn.
Giun c di 2 5mm x 0,1 0,2mm, u thng, ui cong v gp v pha bng. C quan sinh dc
c gm c 1 tinh hon v 1 ng dn tinh. Cui ui c 1 gai sinh dc nh ra ngoi.
Giun ci di 9 12mm x 0,3 0,5mm, u nhn, ui di v nhn v thng, l sinh dc nm 1/3
trc ca thn. C quan sinh dc gm c 2 bung trng hnh ng, hai ng dn trng v hai t cung.


1.8. Giun ch Bch tuyt
Vit Nam thng gp 2 loi giun ch k sinh ngi:
Wuchereria bancrofti.
Brugia malayi.
a) Wuchereria bancrofti
Giun ch trng thnh ging nh si t mu trng sa.
Giun c di khong 3cm, chiu ngang 0,1mm.
Giun ci di khong 8 10cm, chiu ngang 0,25mm.
Giun c v ci thng sng cun vo nhau trong h bch huyt.
Giun ci ra u trng, u trng xut hin trong mu ngoi vi theo chu k m hoc ty theo loi.
b) Brugia malayi
Con trng thnh ging W. bancrofti v hnh thi, nhng mnh v ngn hn.
Giun ci di 4,3 5,5cm.
Giun c di 1,3 2,3cm.
2. HNH TH U TRNG GIUN
2.1. u trng giun mc
Thng ta ch thy u trng ca giun mc trong phn khi phn c gi li mt thi gian trc khi
quan st trng c thi gian n thnh u trng.
Trong phn u trng khng bao gi i qu giai on filariform, khng bao gi n c giai on
trng thnh.

Cc c im phn bit u trng giun mc v u trng giun ln
a) u trng giai on 1 (Rhabditiform)
Kch thc: 250 x 17m.
Ming m, bao ming di v hp.
Thc qun phnh ra pha sau thnh
hnh qu l.
ui thon di v nhn.
b) u trng giai on 3 (Filariform)
Kch thc: di 500 600m.
Ming ng.
Thc qun hnh ng.
ui thon di v nhn.

2.2. u trng giun ln (Strongyloides stercorslis)
u trng thng thy nht trong cc u trng ca giun.
Khi cy phn, c th thy u trng giai on 1, giai on 3 v giun trng thnh.
a) u trng giai on 1 (Rhabditiform)
Kch thc: 200 380 x 16 20m.
Phn sau thc qun phnh ra thnh hnh
qu l.
Ming h, bao ming ngn.
ui thon di v nhn.
b) u trng giai on 3 (Filariform)
Kch thc: 500 630 x 16m.
Thc qun hnh ng.
ui ch hai.

c im u trng giun mc u trng giun ln
u trng
Ming H, bao ming di H, bao ming ngn
ui Nhn Nhn

2.4. u trng giun ch

a) Wuchereria bancrofti
Kch thc: 161 305m.
Thn c nhng ch un cong mm mi v u.
Bao bc ngoi di hn thn nhiu.
Nhn dinh dng nh, t, tch bit nhau r rng, khng i ti cui ui.
ui nhn, thon u t t, khng c nhn.
giai on
1
Rhabditiform
Thc qun Phnh hnh qu l Phnh hnh qu l
Di ng Chm Nhanh
Hnh dng Mp Mnh
Hin din
trong phn
Sau 24 48 gi C ngay trong phn
u trng
giai on
3
Filariform
Ming Kn Kn
ui Nhn Ch hai
Thc qun Hnh tr chim 1/4 thn Hnh tr chim 1/2 thn
2.3. u trng giun xon (Trichinella spiralis)

u trng cun thnh hnh xon c trong nang c
vn ca ngi v ng vt.
Nang giun xon c hnh bu dc di 200
400m.
u trng c kch thc
90 100m x 60m. Khi mi vo c th, u trng
c hnh gy v c mng bao. Sau khi nhim 21
30 ngy, u trng c mng bao bc bn ngoi.


b) Brugia malayi
Kch thc: 180 230m.
Thn c nhng ch un cong khng u, ui xon.
Bao bc ngoi di hn thn t.
Nhn dinh dng nh, nhiu, st nhau, khng r.
ui nhn, c 2 nhn.

c im u trng Wuchereria bancrofti u trng Brugia malayi
Hnh th Qun t Qun nhiu
Kch thc 8 x (220 300)m 5 x (160 220)m
Khong u Chiu di gn bng chiu ngang Chiu di gp i chiu ngang
ui Thng Tn cng c 2 nhn
Nhn bn trong R v khng lan n tn ui Khng r v lan n tn ui
Thi im xut hin
mu ngoi vi
Khong 21 gi 2 gi sng Khong 24 4 gi sng
Ni k sinh Mu ngoi vi v ni c dch bch
huyt tch ly
Mu ngoi vi

CU HI LNG GI
1. Anh (ch) da vo cc c im no nh danh giun, sn?
2. M t hnh th ca giun. Giun c khc giun ci nhng im no?
3. M t cu to ca giun.
4. Anh (ch) cho bit nhng im khc bit gia giun kim v giun mc.
5. So snh hnh th ca u trng giun mc vi giun ln.
6. Da vo c im no nhn bit l u trng giun ch h bch huyt?
7. Anh (ch) cho bit nhng im khc bit gia u trng Wuchereria bancrofti v Brugia malayi.

Bi 25
NHN DNG CC LOI SN L

xc nh bnh sn l, ngi ta da vo s tm trng sn trong phn bng cch soi mu phn di
knh hin vi.
Sn l trng thnh c th c nhn thy bng mt thng v ch c thy trong phn sau khi bnh
nhn dng thuc ty sn.
1. C IM CHUNG CA SN L


Thn dp, hnh l cy, khng c t, kch thc t vi milimt n vi centimt.
C 2 a ht, mt ming v mt bng.
C quan tiu ha gm c ming nm y a ht ming, thng vi thc qun v rut. Rut c 2
nhnh bt kn, khng c hu mn.
Sn l lng tnh.
B phn sinh dc c gm c hai tinh hon c hnh khi hoc chia thy hoc thnh nhnh. Mi tinh
hon c mt ng dn tinh, sau mt on, 2 ng ny nhp li thnh 1 ng duy nht i ti c quan giao hp
c, gi l dng vt, m ra bng, gn a ht bng. Dng vt c th nm trong mt ti gi l ti dng
vt.
B phn sinh dc ci gm c mt bung trng, mt u thng vi hai tuyn nui dng bung trng
hai bn thn v u kia thng vi t cung. T cung l mt ng di, khc khuu, cha trng v i ti l
nm cnh l sinh dc.
2. SN L RUT (Fasciolopis buski)
Thn dp, dy, mu nu sm hay xm.
Kch thc: 20 70m x 8 20m.
u khng c phn nh ln.
Trn thn c nhng gai nh xp thnh hng, nhiu nht l gn a ht bng.
a ht bng ln hn a ht ming.
Rut c 2 nhnh i ti tn cui ui.
Tinh hon chia nhnh rt nhiu chim ht c phn gia v phn sau ca thn.
Bung trng cng chia nhnh.
T cung nm pha trc ca thn, cha nhiu trng.
L sinh dc trc a ht bng.

3. SN L GAN LN (Fasciola hepatica)
Di t 2 4cm, thn dp, dy.
u nh nhn, c th hnh nn u.
Pha trc thn c phn nh ln hnh nn, pha sau thn dp v trn.
a ht ming ln hn a ht bng.
Rut di n tn cui thn.
L sinh dc nm trc a ht bng.
C ti dng vt, tinh hon nm sau bung trng.
Tinh hon, bung trng v rut u phn nhnh rt nhiu.


4. SN L GAN NH (Clonorchis sinensis)
Mu trng c hoc mu nht.
Kch thc: 10 25mm x 3 4mm.
C th khng ph gai.
Thc qun kh di, 2 nhnh manh trng kn i n tn cui ui.
a ht ming ln hn a ht bng.
2 tinh hon phn nhnh, phn b ci trn ci di, nm sau bung trng.
Bung trng v rut khng phn nhnh.
T cung gia bung trng v a ht bng.
L sinh dc nm trc a ht bng. Khng c ti dng vt.


5. SN L GAN NH (Opisthorchis felineus v Opisthorchis viverrini)
Thn c hnh nh 1 ci chai, c 2 a ht, mt u (phn ming) v mt bng.
Tinh hon nm di bung trng, tinh hon phn thy, 2 ci chch nhau .
T cung gia bung trng v a ht.

6. SN L PHI (Paragonimus westermani)
Thn dy mt trn li, mt bng dp gn ging
7. SN MNG

Sn c mu trng, v sn nhm.
Di 1,3cm, rng 1mm.
C a ht ming v a ht bng dng bm.
Thn cun li nh lng mng, gia thn c rnh su mang sn ci.
nh ht c ph, mu nu .
Kch thc: 7 12mm x 4 6mm.
Trn thn c nhiu gai nh.
a ht ming v a ht bng c kch thc
bng nhau.
Rut mnh mai, ngon ngoo, khng phn
nhnh, i ti cui thn.
Bung trng to chia thnh thy 2 bn.
Tinh hon phn thy nm sau bung trng.
L sinh dc nm sau a ht bng.

Hai manh trng nhp li pha sau to thnh on manh trng duy nht.
Sn c c 4 5 tinh hon.
Sn ci mnh v di hn trung bnh 2,2cm, t cung di cha 20 30 trng.

CU HI LNG GI
1. M t hnh th v cu to ca sn l.
2. Anh (ch) cho bit nhng im khc bit gia sn l rut v sn l gan ln.
3. So snh hnh th ca Clonorchis sinensis v Opisthorchis viverrini.
4. M t hnh th sn mng trng thnh.

Bi 26
NHN DNG CC LOI SN DI (SN DY)


Thng thng, vic chn on bnh sn di da vo s tm thy nhng t sn trong phn hoc dnh
trn qun o bnh nhn, tr trng hp Diphyllobothrium latum, sn thng trng v Hymenolepis, trng
thot theo phn ra ngoi do t sn thng b tiu nt trong rut.
Do , ta cn phi quan st phn bng mt trc. Ty t sn tm thy, ta s nh loi da trn nhng
c im ca chng.
1. C IM CHUNG CA SN DI
Mt u tht nh thng c a ht v c mc hoc rnh ht.
Mt c nh, khng c c quan r rt, t y sn sinh nhng t non bng cch ny chi.
Thn gm mt chui cc t ni vi nhau to thnh si dy, nhng t cng xa u th cng ln v
cng gi.

nhng t cui cng, cc c quan sinh dc teo li, ch cn li t cung phn nhnh, cha nhiu trng.
2. SN DI HEO (Taenia solium)
2.1. Sn trng thnh
Chiu di thn: 2 8m, c khong 800 1000 t.
u hnh cu, ng knh khong 1mm, c 4 a ht trn v mt chy nh cao mang 2 vng mc.
t c ngn v mnh.
Nhng t gn u c chiu ngang ln hn chiu di, cng v pha ui, t sn cng di ra, nhng
t cui chiu ngang ch bng 1 na chiu di.
t trng thnh c chiu ngang bng chiu di cha c quan sinh dc c v ci. L t cung mt
Sn di lng tnh.
C quan sinh dc c gm nhiu tinh hon
di dng ht nh, ni lin vi nhau bng ng dn
tinh nh. Nhng ng dn tinh nh ny vo mt
ng dn tinh chung, ng ny thng i ra mt bn
ca t ti l sinh dc.
C quan sinh dc ci gm c 1 hay 2 bung
trng, 1 ng dn trng, tuyn dinh dng, t cung
v m o. m o v ng dn tinh chung vo
mt ch gi l l sinh dc.

bn.
t gi mang trng:
+ Chiu di ln hn chiu rng (1 2cm x 0,5 0,7cm).
+ 1 l sinh dc mt bn cnh, xen k tng i u 2 bn.
+ T cung l mt ng chy theo chiu dc gia t, phn nhnh ra 2 bn, cha y trng.
+ S nhnh t cung mt bn t, di 13 nhnh.
+ Nhng t gi cui thn thng rng thnh tng on ngn 5 6 t dnh nhau v c thi ra
ngoi theo phn.
+ Khng di ng.

2.2. Nang
u trng
sn di heo
(Cysticercus
cellulosae)
L mt
bc trn
hay bu
dc.
C
kch
thc
(0,7
3. SN DI B (Taenia saginata)
Chiu di thn: 2 8m, c th n 10m.
u sn hnh tri l ng knh 1 2mm, c 4 a ht trn, khng c chy v mc.
t gi mang trng:
+ C chiu di gp 2 3 ln chiu ngang (1 2cm x 0,5 0,7cm).
+ L sinh dc mt bn cnh, xen k khng u.
+ T cung l mt ng chy theo chiu dc chnh gia, phn nhnh ra 2 bn, cha y trng.
+ S nhnh t cung mt bn nhiu hn 13 nhnh.
+ t sn thng t b ra ngoi ring r, ri trn qun o hoc ging chiu, t khi b thi ra ngoi ra
ngoi theo phn.

4. SN DI C (Diphyllobothrium latum)
C thn di t 3 10m, khong 3000 t.
C mu trng ng hoc xm khi.
u sn hnh thun, kch thc 2 3mm x 0,7mm, khng c a ht, m c 2 rnh ht.
t trng thnh c hnh thang, chiu ngang ln hn chiu di. L sinh dc gia t, l t cung gn
0,8cm) x
1,5cm.
Bn
trong cha
dch v u
u trng sn.
u c
4 a ht
v 2
hng
mc.
l sinh dc.
t sn mang trng: chiu ngang ln hn chiu di (3 x 11mm), c 1 l sinh dc chnh gia. T
cung l 1 ng un thnh nhng vng cun nm ngay gia t sn, cha y trng.

5. SN DI CH (Dipilidium caninum)

6. SN DI (Hymenolepis nana)
Sn nh, ch di t 1 2cm, khong 200 t sn.
u sn hnh cu, ng knh 0,25mm, c 4 a ht hnh chn, c 4 a ht v mt chy c th nh ra
C thn di 10 50cm. C
khong 60 175 t, mu hng nht.
u c 4 a ht hnh chn, chy
mang 3 7 hng mc, c th nh ra
hoc tht vo.
Nhng t gn u rt ngn,
nhng t gia hnh dng nh thng
ru.
t sn mang trng c hnh ht
da.

t sn di hn rng
(0,8 1cm x 0,3cm), hnh ging ht da, 2 l sinh dc 2 bn, t cung gm nhng ti ng
trng (27mm x 12mm).

hay tht vo vi mt hng mc.
t sn trng thnh hnh thang.
C quan sinh dc c gm c 3 dch hon trn, 1 ng dn tinh tn cng bng ti dng vt. Bung
trng c 2 thy nm gia t, t cung hnh ti nm ngang, l sinh dc nm cng mt bn.
t sn mang trng c mt t cung hnh ti cha 80 180 trng.

7. SN DI (Hymenolepis diminuta)
Di 4cm.
u sn c 4 a ht, chy nh u ngn, khng c mc.
t sn mang trng nh chng vi mm, chiu rng ln hn chiu di, l sinh dc mt bn, t cung
hnh ti nm ngang.
Mt v bc ngoi c vn, dy, khng nhn, gm nhiu phin ng tm.
Mt mng sinh mm, c nhiu nhn, nhiu t bo cha cht d tr.
Nhiu nang sinh mm (brood capsules) pht sinh t mng sinh mm, cng cu to nh mng sinh
8. SN DI CH (Echinococcus granulosus)
8.1. Sn trng thnh
Sn nh nht, kch thc: 2,5 9mm.
u sn hnh trn c 2 vng mc v 4 b phn
bm.
Ba t u: b phn sinh dc cha pht trin
hon ton, t cui pht trin y .
t sn c 12 15 nhnh t cung.
Trng tng t cc loi khc, kch thc 30
38m.
8.2. Nang thng (Hydatid cyst)
Hnh trn, ng knh 1 7cm, gm 4 phn t
ngoi vo trong.

mm v ny chi cho nhiu u sn tht vo trong, c mc.
Mt dung dch trong, khng c li khi b h nng hay trn vi acid.

8.3. Ct sn (Hydatid sand)
Cc u sn nhiu khi ri rng xung to thnh mt lp di y nang thng ging nh nhng ht ct.
Cc u sn ny c th tht vo hay nh ra.

CU HI LNG GI
1. M t hnh th v cu to ca sn di.
2. Anh (ch) cho bit nhng im khc bit gia sn di heo v b.
3. M t go u trng sn di heo.
4. So snh u v t trng thnh ca Taenia solium v Diphyllobothrium latum.
5. Phn bit u ca Taenia solium vi u ca Dipilidium caninum.

Bi 27
HNH TH TRNG GIUN, SN THNG GP


1. C IM CHUNG CA TRNG GIUN, SN
1.1. Hnh th
Trng giun, sn c nhiu hnh th khc nhau: hnh trn, thon di hoc hnh thoi, cn i hoc khng cn
i, hoc lp 1 gc. Trng sn l c np nh 1 u, u i din c 1 nm hoc v dy ln. Trng sn
mng c gai.
1.2. Kch thc
Trng giun, sn c kch thc to nh nhau ty theo tng loi trng. Trng sn l gan nh c kch thc
nh nht, trng sn l gan ln c kch thc ln nht. Khi so snh kch thc, phi quan st cng mt
phng i.
1.3. Mu
Thng th trng c mu vng ca phn nh trng giun a, giun tc, trng sn l gan nh, sn l phi,
sn l gan ln, trng sn l rut. C trng khng mu nh trng giun kim.
1.4. Cu to
Gm 2 phn:
V: c loi v dy, gm nhiu lp nh trng giun a, giun tc, trng sn di, c loi v mng ch c
1 lp nh trng giun kim, giun mc.
Nhn: nhn ca trng thay i ty theo giai on pht trin:
+ Trng mi sinh ra: nhn l mt khi, cha pht trin: trng giun a, trng giun tc.
+ Trng sinh ra mt vi ngy: nhn phn chia thnh nhiu mi hoc hnh thnh u trng bn trong nh
trng giun mc, trng giun kim.
+ Bn trong trng c mc: trng sn di.


2. HNH TH CA CC LOI TRNG GIUN
2.1. Trng giun a (Ascaris lumbricoides)
Trng giun a c 3 loi:
a) Trng th tinh (trng chc)
C hnh bu dc hoc hi trn, rt cn i.
Kch thc khong 45 75m x 35 50m.
Mu vng.
V dy, gm 3 lp ng tm:
+ Ngoi cng l lp albumin dy u, x x.
+ Lp gia dy, nhn v trong sut c cu to bi glycogen.
+ V mt lp v trong cng l mng dinh dng cu to bi lipid.
Bn trong trng l phi bo chc, gn thnh 1 khi, cha phn chia khi trng mi c ra.

b) Trng khng th tinh (trng lp)
C hnh bu dc di v hp hn, hnh dng k d.
Kch thc t 88 94m x 39 44m.
V ch c 2 lp mng, khng c lp mng dinh
C hnh dng bt thng do lp albumin b trc lm mt lp ngoi sn si, v trng tr nn nhn.
Trng ch c mt v dy v nhn v cc vn ng tm.
Gp trng th tinh hay khng th tinh.

2.3. Trng giun mc (Ancylostoma duodenale/Necator americanus)
2.4. Trng giun kim (Enterobius vermicularis)
dng.
Bn trong trng khng c phi, ch c nhng ht trn
khng u, rt chit quang.
c) Trng mt v

2.2. Trng giun tc (Trichuris trichiura)
Hnh bu dc, hi di, rt i xng, hai u c
2 nt nhy.
Kch thc: 50 65 x 20 29m.
Trng c mu vng m ng sang nu.
V dy, nhn gm 3 lp.
Trng mi c ra c phi bo, cha phn
chia.

Kh phn bit trng ca 2 loi giun ny bng
cch quan st.
Hnh bu dc, hi di, rt i xng.
Kch thc: 50 70m x 40m.
V mng, nhn trong sut.
Trng mi c ra c t 2, 4 hoc 8 phi
bo.
Trng giun mc ra ngoi cnh thng sau
24 gi n thnh u trng.

Hnh bu dc, thon di, khng cn i,
lp 1 bn.
Kch thc: 50 60m x 20 30m.
Trng khng c mu, trong sut.
Ta khng thy trng ca giun ln trong phn nhng c th thy trong cc bnh phm ht t rut ra.
Trng giun ln bao gi cng c phi thai, rt ging trng giun mc, nhng kch thc nh hn (50 x
30m).
3. HNH TH CA CC LOI TRNG SN L
3.1. Trng sn l rut (Fasciolopsis buski) v sn l gan ln (Fasciola hepatica)

Hnh bu dc.
Kch thc 160 150m x 85 95m.
Trng mi c thi ra ngoi cnh c mu vng nht.
V mng, nhn, c np.
Bn trong cha phi bo.
V mng.
Trng cha u trng bn trong.
2.5. Giun ln

3.2. Trng sn l gan nh (Clonorchis sinensis)
Hnh bu dc, nh, ch gia np v phn cn li
nh ra r.
Kch thc 27m x 18m.
Trng thng c mu vng.
V c 2 lp, nhn v mng, np li rt d thy v
mt nm pha i din np.
Trng c phi ngay t lc mi sinh.

3.3. Trng sn l gan nh (Opisthorchis felineus)
Trng nh, hp, kch thc 30 x 11m i xng, c phi thai lch.
4.1. Trng sn di heo v sn di b (Taenia solium v Taenia saginata)
3.4. Trng sn l phi (Paragonimus westermani)
Hnh bu dc, i xng.
Kch thc: 80 120m x 45 60m.
Trng c mu vng nht hoc mu nu sm.
Np dp, khng theo ng vng cung ca trng.
V mng, nhn, dy ln pha i din vi np.
4. HNH TH CC LOI TRNG SN DI

Hnh trn.
ng knh khong 40m.
Mu nu sm.
V dy, c 2 lp, c vch hng tm t
trong ra ngoi.
Cha mt phi c 6 mc.

4.2. Sn di c (Diphyllobothrium latum)
Hnh bu dc.
70 x 45m.
Mu nu.
C np nh.
Khi mi sinh ra ch c cc t bo phi.

4.3. Sn di ch (Dipilidium caninum)
Hnh trn.
Khng mu sc.
V dy.
Trng c cha trong mt ti sinh trng.
Bn trong c phi sn.
4.4. Sn di (Hymenolepis nana)

Hnh bu dc.
ng knh khong 40m.
C v dy c 2 mng: mng ngoi mng,
mng trong dy, mi cc c mt chm nh v
t xut pht nhiu si tua nh.

5. HNH TH TRNG SN MNG
5.1. Trng sn mng (Schistosoma haematobium)
Hnh thun.
Di 120 140m.
V xm v chit quang.
C gai nh mt cc.
Trng cha u trng c lng t di ng.
Thy trong nc tiu.

5.2. Trng sn mng (Schistosoma mansoni)
Hnh thun.
Di 120 140m.
Mu nu sng.
C gai mt bn.
Cha u trng c lng t di ng bn trong.
Thy trong phn.

Bn trong trng c mt phi thai
6 mc.

4.5. Sn di (Hymenolepis diminuta)
Hnh cu hoc hi bu dc.
ng knh khong 75m.
V khng mu sc.
Mng ngoi dy.
Khng c cc si tua.


5.4. Trng sn mng (Schistosoma mekongi)
C hnh dng tng t nh trng S. japonicum nhng nh hn, di 55 65m.
5.3. Trng sn mng (Schistosoma japonicum)
Hi trn.
Kch thc: 70m.
Mu nu xm.
V nhn.
C gai nh bn, cha u trng c lng t di ng.
Thy trong phn.



CU HI LNG GI
1. Anh (ch) da vo nhng c im no nh danh trng giun, sn?
2. M t trng giun a chc v lp.
3. Trng giun tc khc trng giun a nhiu im, nhng c mt im ging nhau v cu to. Anh (ch)
cho bit im ging nhau .
4. So snh trng giun mc v trng giun kim.
5. Cho bit nhng c im chung ca trng sn l.
6. M t trng Taenia sp. Anh (ch) c th nhn dng nhm gia trng Taenia vi trng ca giun, sn no
khc?
7. Nu nhng im khc nhau ca trng gia 3 loi sn mng (Schistosoma haematobium, Schistosoma
mansoni v Schistosoma japonicum).
8. Cho bit tn ca trng giun, sn nh nht v ln nht c th gp trong phn.


Bi 28
N BO NG RUT

1. I CNG
n bo l nhng sinh vt m c th ch c cu to bi 1 t bo, c chia lm 4 lp ty theo cch
sinh sn v c quan chuyn ng:
1. Lp trng chn gi (Rhizopoda)
2. Lp trng roi (Flagellata)
3. Lp trng lng (Ciliata)
4. Lp trng bo t (Sporozoa)
Ngoi tr cc n bo lp trng lng v cc trng nang ca trng bo t, kch thc ca cc n bo
thng trong khong t 5m n 30m, v vy, vi vt knh x10, trng chng ging mt im phn chiu
nh sng.
+ Trong phn lng: thng ch thy th hot ng ca cc n bo k sinh trong rut.
+ Trong phn cng: ch thy bo nang ca n bo.
+ Trong phn nt, nho: c th thy c th hot ng v bo nang, th hot ng thy ch c m,
cht nhy v mu.
Th hot ng b thoi ha rt nhanh nn cn phi xem sm, cng sm cng tt. Tt nht l xem phn
qua knh hin vi ngay khi bnh nhn va i tin xong.
+ Nu nhn c nhiu mu phn, ta phi xem theo th t u tin: phn lng trc nht, phn c
sau cng (v bo nang ca n bo vn thy r sau nhiu ngy).
+ S quan st trc tip qua knh hin vi v khng nhum trc gip ta thy th hot ng nh s di
ng ca chng v nh s phn chiu nh sng.
Ta c th c lng kch thc, tnh cht ca c quan chuyn ng v s n (thc bo) hng cu ca th
hot ng.
2. TH HOT NG
nh danh th hot ng ca n bo thng da vo cc yu t sau y:
Kch thc
T bo cht
Chn gi
Nhn
Ngoi t bo cht
Ni t bo cht
Khng bo
Cc n th
Mng nhn: th nhim sc
Nhn th
Roi
Mng ln sng.

2.1. Quan st trc tip
a) Entamoeba histolytica
Hnh dng khng nht nh khi chuyn ng, c hnh trn khi ng yn.
Kch thc: 12 40m.
Di chuyn kh nhanh, c nh hng.
Ngoi t bo cht trong; ni t bo cht c ht, cha khng bo nh.
Chn gi di v trong.
Nhn trn, khng thy r khi amp cn sng, ng knh khong 5m.
Trn mng nhn c nhng ht nhim sc nh xp u n.
Nhn th nh ging nh mt chm gia nhn.
Trong phn lng, c th gp c 2 dng: gy bnh v khng gy bnh.
+ Dng gy bnh: kch thc 20 35m, n hng cu.
+ Dng khng gy bnh: kch thc 12 20m, khng bo cha vi trng v cc mnh vn khc.

b) Entamoeba coli
Kch thc: 20 40m.
Hnh dng khng u, c th bu dc hoc di, rt chit quang.
t di ng, hoc di chuyn chm chp, khng nh hng, gi tc to, ngn.
T bo cht c ht, ngoi v ni t bo cht khng r.
C nhiu khng bo cha vi trng,
nm, mnh vn t bo nhng khng bao gi
c hng cu.
Nhn thy r khi cn sng, c hnh
trn, trn mng nhn c nhng ht nhim
sc nh xp khng ng u.
Nhn th kh to, nm lch tm

c) Entamoeba hartmani
Kch thc: 3 7m, ti a l 15m.
T bo cht cha khng bo nh
Nhng c im khc ging E.
histolytica, nhng khng bao gi n hng
cu.

d) Endolimax nana
g) Giardia lamblia
Thn hnh qu l, mt bng phng, mt lng g.
Kch thc 10 20 x 6 10m.
Chuyn ng nh l ri, c nh hng.
C trc sng lng dc theo thn.
a ht nm mt bng.
C hai nhn, mi nhn c mt nhn th to.
T gc roi u truc, 4 i roi xut pht i v pha sau thn.
Kch thc: 6 10m.
C nhiu gi tc trn, nh, trong, di
chuyn chm chp, khng nh hng.
T bo cht c nhiu ht v cha
nhiu khng bo.
Khng bo cha nhiu cht khc
nhau, ch yu l vi trng.

Nhn khng r khi amp cn sng.
Nhn th ging nh mt git mc, c th nhn thy sau khi nhum vi iod.
e) Iodamoeba butschlii
Kch thc: 10 15m.
Hnh dng ging chic l.
Di chuyn rt chm, gi tc trong,
trn ging nh ngn tay.
T bo cht c nhiu khng bo,
cha vi trng, nm, tinh bt,
Nhn kh to (4 6m), mng nhn
mn.

Nhn th to hnh trn hoc bu dc, thng gia nhn.
f) Dientamoeba fragilis
Hnh th: a dng.
Kch thc: 6 15m.
Rt di ng nhng nhanh chng
mt tnh di ng sau khi c thi ra
ngoi.
Ngoi t bo cht trong, c nhiu
khng bo.

Khng bo cha vi trng, nm, tinh bt.
C 1 hoc 2 nhn, c th nhn thy sau khi nhum vi iod.
Nhn th chia lm 4 6 ht, rt kh thy mng nhn.

h) Trichomonas hominis
Hnh qu l hay hnh cu.
Kch thc: 10 15 x 7 10m.
Rt di ng, chuyn ng khng nh hng.
Thn c ch lm.
C mt nhn to vi nhiu hch nhn pha
u.
3 5 roi t do pha trc v c 1 roi dnh
vo thn to thnh mt mng ln sng di ti
cui thn.
Trc sng lng chy dc thn.
T bo cht cha nhiu khng bo.
T. hominis khng c bo nang.

i) Chilomastix mesnili
Hnh qu l, thn nh b vn v nhn
v pha sau.
Kch thc: 10 15 x 5 6m.
Thn c mt rnh xon c.
Khng c mng ln sng.
C 3 roi di, hung v pha truc.
C 1 nhn.
Di chuyn theo ng vng l xo v mt
hng nht nh.

k) Enteromonas hominis
Hnh qu l.
Kch thc: 4 10 x 3 6m.
C 3 roi di, hung v pha truc v 1 roi
hng v pha sau.
2.2. Sau khi nhum trichome
a) Entamoeba histolytica
C 1 nhn to.
Di chuyn theo ng vng l xo v mt
hng nht nh.

l) Retortomonas intestinalis
Hnh qu l.
Kch thc: 5 17 x 2 3m.
C 2 roi di, hng v pha trc.
C 1 nhn.
m) Trichomonas vaginalis

Hnh cu hay hnh qu l.
Kch thc: 15 30 x 7 10m.
C mt nhn to.
C trc sng lng chy dc thn.
4 roi xut pht t mt gc roi hng ra pha
trc v 1 roi hng v pha sau, dnh vo thn to
thnh mt mng gn sng ngn.
T bo cht cha nhiu ht v nhiu khng
bo.
Khng c th bo nang.

n) Balantidium coli
Thn hnh bu dc c lng t bao quanh.
Kch thc: 50 100m.
C 1 ch lm mt u, gi l ming, c
lng t di.
Hu mn nm i din vi ming.
T bo cht lm tm ht.
C mt nhn to hnh ht u, mt nhn nh
hnh trn nm cnh ch lm ca nhn to.
Nhiu khng bo tiu ha v bi tit.
Chuyn ng nhanh v u.

Hnh trn, i khi hi di.
ng knh: 10 22m.
Chn gi n mu nht.
T bo cht c ht mn.
Khng bo nh v khng thy r.
Nhn c ng knh t 3 5m.
Mng nhn mnh, u.
Nhn th gia nhn.
b) Entamoeba hartmani
ng knh 3 14m, trung bnh: 6m.
C nhiu chn gi a ra nhiu hng.
T bo cht c khng bo nh v th vi.
Nhn th c th gia nhn hoc lch tm.
c) Entamoeba coli
Hnh trn, i khi hi di.
ng knh t 10 35m.
Chn gi t, ngn.
T bo cht c ht.
Khng bo to v cha y vi trng, nm, mnh t bo.
Mng nhn ch dy, ch mng, khng u.
Nhn th to, lch tm.
d) Endolimax nana
Mng nhn mnh.
T bo cht cha nhiu khng bo nh, r.
Nhn th trn, gia nhn c th bu dc, lch tm.
e) Iodamoeba butschlii
Trn, bu dc hoc di.
ng knh trung bnh 10,2m.
Chn gi trong.
T bo cht cha nhiu khng bo y vi trng, nm, tinh bt.
Mng nhn mnh.
Nhn th trn thng gia nhn.
3. TH BO NANG
Bo nang l th c tnh khng vi nhng yu t bt li trong mi trng v l th ly nhim ca cc
n bo thuc lp chn gi, trng roi v trng lng.
Th bo nang thng trn, c kch thc nh, bt ng, c th cha mt nhn hay nhiu nhn.

Th bo nang ca n bo ng rut c nh danh nh cc c im nh sau:
Hnh th
Kch thc
Cc c quan trong t bo cht
Nhn
Mng nhn: th nhim sc
Nhn th.
3.1. Entamoeba histolytica
Hnh trn, chit quang.
V dy gm 2 lp.
Kch thc: loi ln (10 15m) v loi nh (di 10m).
C t 1 4 nhn.
C nhng ht nhim sc nh phn b u n trn mng nhn.
Bo nang tr (1 2 nhn) c nhiu c cu hnh que c 2 u trn, chit quang gi l thi chromatoid,
i khi c khng bo to.
Bo nang c 4 nhn: khng bo v cc th chromatoid bin mt.


3.2. Entamoeba hartmani
Hnh trn, i khi bu dc.
Kch thc: 4 8m.
C t 1 4 nhn.
Vch dy

3.3. Entamoeba coli
Hnh trn hoc hi bu dc.
Kch thc: 12 25m.
C 1 8 nhn (thng gp 8 nhn).
Nhn hnh trn.
C nhng ht nhim sc th phn b khng u trn
mng nhn.
Nhn th ging mt vt nho, lch v mt bn.
C nhng c cu hnh que, mnh, di, c 2 u
nhn, gi l thi chromatoid.
i khi c khng bo to, thng gp nhng bo
nang tr (1 2 nhn).

Bo nang tr (1 2 nhn) c nhiu c cu hnh que c 2 u nhn, chit quang gi l thi
chromatoid, i khi c khng bo to. Nhng c cu ny dn dn thu nh li v bin mt khi bo
nang pht trin n giai on gi.
3.4. Endolimax nana
Hnh bu dc.
Kch thc: 7 12m.
Vch mng, t bo cht trong.
C 4 nhn trng nh 4 vt nhe, khng r.
Nhn th ln, bao bc bi mt qung sng.

3.5. Iodamoeba butschlii
Hnh trn hoc bu dc.
Kch thc: 8 15m.
Vch kh dy.

T bo cht trong v chit quang.
C mt khng bo cha glycogen.
Nhn cha mt nhn th nm lch tm.
Nhn th trn, bu dc, ln, bao bc bi mt qung
sng.

3.6. Giardia lamblia
Hnh bu dc.
Kch thc: 9 13m.
C mt trc sng thn chnh gia bo nang.
C t 2 4 nhn v c th c vi roi xp li ging
hnh ch S trong bo nang.

3.7. Chilomastix meslini
C hnh qu l, rt chit quang.
Kch thc: 6 10m.
Vch dy.
C mt nhn, to, b y v mt bn.
Trong bo nang c cc vt dc.

3.8. Enteromonas hominis
Kch thc 6 8m.
C 2 4 nhn, ghp thnh i 2 cc.
3.9. Retortomonas intestinalis

Hnh qu l, nh nht trong cc bo nang k sinh
ngi.
Kch thc: 4 7m.
Khng mu.
Vch kh dy.
Khng thy r c cu bn trong vch.

3.10. Balantidium coli
Hnh trn hoc trng.
Kch thc: 50 80m.
Vch dy c 2 lp.
C 1 nhn ln.
Nhiu khng bo v nhiu th chit quang.

4.1. Cryptosporidium sp

4.2. Isospora belli

Trng nang c hnh xoan, di, c mt u nhn hn u kia.
Kch thc: 25 33 x 12 16m.
V nhn, bn trong c mt bo t nguyn bo gia.
Sau khi ra ngoi cnh, bo t nguyn bo ny s pht trin ln thnh 2 bo t nang, mi bo t nang
c 4 thoa trng.
4. TRNG NANG

Trng nang c hnh cu.
ng knh 5 6m.
Vch mng.
T bo cht c ht mn, vi mt m en gia hoc
gc.
C 4 thoa trng hnh lim hay ngn tay.
C th nhn thy nhn gia thoa trng.

4.3. Sarcocystis sp
Bo t nang c hnh trng, rt chit quang.

Cc th hot ng rt chng b thoi ha nn vic nh danh n bo k sinh trong rut phn nhiu nh
tm thy bo nang. Khc vi th hot ng cn phi xem ngay, cc mu phn nghi c th bo nang c th
c gi li hoc nhit thng, hoc trong t lnh, cc bo nang c thi gian ny ny n
trong trng hp nghi ng, nn dnh mu phn trong t lnh, 24 gi sau xem li.
Bo nang cha n :
+ Rt kh xc nh.
+ Nhn cng ging nhn ca th hot ng v bo nang cng loi.
+ Khc vi bo nang ny n ch:
* S nhn t hn nhng nhn to hn.
* Khng bo c glycogen nhum mu nu sm nhiu hay t vi Iod.
Nhng vt sau y c th lm vi bo nang:
+ Ht m: kch thc khng u, c th nhum bng thuc nhum c bit.
+ Tinh bt: nhum mu tm vi Iod.
+ Bch cu: nhum Iod, cc cu to ca nhn khng thy r.
CU HI LNG GI
1. Anh (ch) da vo nhng c im no nh danh th hot ng ca n bo?
2. K tn cc loi amp thng gp ngi.
3. M t th hot ng ca Entamoeba histolytica.
4. Nu nhng im khc bit gia th hot ng ca Entamoeba histolytica v Entamoeba coli.
5. M t th hot ng ca Giardia lamblia.
6. Anh (ch) nhn dng th bo nang n bo da vo yu t no?
7. C g khc nhau gia bo nang tr ca amp v bo nang hon chnh; gia bo nang ca Entamoeba
histolytica v Entamoeba coli?
8. M t hnh th ca Cryptosporidium.
9. Khi quan st tiu bn phn, anh (ch) c th gp nhng kh khn no?
Kch thc: 14 15 x 9 10m.
C 4 thoa trng c dng hnh lim.

4.4. Cyclospora sp
Hnh cu.
ng knh: 9 10m.
V c 2 lp mng: mng ngoi cng rt dy.
Bn trong c nhiu ht rt chit quang ni ln trng
ging nh tri du.
5. IU CN BIT TRONG KHI TM BO NANG


Bi 29
NHNG VT TH C TH TM THY TRONG PHN

1. T BO THC VT C TINH BT
Kch thc: 50 100m.
Hnh trn hay bu dc, ng vin xung quanh bao gi cng mo m, khng phng.
Bn trong l nhng ht tinh bt ng st nhau.


Hnh trn.
Kch thc: to, nh khc nhau.
V gi, bn trong rng.

2. SI THT TIU HA
Kch thc: 100 120m.
Hnh bu dc hoc l hnh ch nht vi nhng
cnh trn
Trong sut hoc c nhng sc ngang.
3. BT KHNG KH, GIT DU



Bi 30
HNH TH K SINH TRNG ST RT NGI

1. I CNG
K sinh trng st rt (KST SR) thuc gii n bo, lp bo t trng, h Plasmodidae, ging
Plasmodium.
y l loi KST sng trong mu c kh nng pht trin nhanh, gy bnh st rt cho ngi v to thnh
nhng v dch ln rt nguy him.
K sinh trng st rt c chu k pht trin tng i phc tp v hnh th a dng nn vic pht hin
phng v tr bnh vn cn ang gp nhiu kh khn hin nay.
C 4 loi Plasmodium c kh nng gy bnh cho ngi, nhng Vit Nam ch c ba loi vi t l
gy nhim nh sau:

2. HNH TH K SINH TRNG ST RT MU NGOI VI
2.1. Hnh th chung ca cc loi KST SR
Hnh th k sinh trng st rt rt phc tp v a dng.
Trn mt tiu bn mu c mang KST SR sau khi nhum Giemsa hay Wright, ta thy:
+ Nhn bt mu thm n tm.
+ T bo cht bt mu xanh nht n xanh tm.
+ Phn khng bt mu l khng bo.
+ Cc ht sc t: en, nu en, nu nh vng.
+ Cc ht c hiu nu, hng nht.
Ty theo hnh thi v s kt hp ca cc yu t trn, ngi ta chia ra cc th ca cc loi k sinh trng
st rt nh sau:
+ Th t dng tr (th nhn) (Early Trophozoite).
+ Th t dng gi (Late Trophozoite).
+ Th phn lit (Schizont).
+ Th giao bo (Gametocyte).

Plasmodium falciparum


c im
Hng cu b k sinh
Tng i bnh thng.
Mt hng cu c th c t 2 3 KST.
S hin din ca cc th Thng thy th nhn v th giao bo trong pht mu ngoi vi, th
t dng gi v phn lit c th gp trong st rt c tnh.
T dng tr
( Th nhn )
Early Trophozoite

C hnh nhn.
T bo cht (TBC) mnh, mu xanh da tri, bao quanh 1 khng
bo ln gia.
Nhn , nm b t bo cht.
i khi thy th nhn 2 nhn.
C th thy hin tng a nhim: nhiu nhn trong 1 hng cu.
Thnh thong gp th kt dnh ngoi vi: KST b dp, dnh vo thnh
hng cu, khng nhn thy khng bo, nhn l 1 chm , nm gia
2 vch ngn.
T dng gi

LateTrophozoite
C dng amp.
T bo cht dy hn c ht sc t nu en.
Nhn to hn.
i khi c m Maurer: ht to nh khng u, nm ri rc trong
hng cu nhim th ny.
Th phn lit
Schizonts

C hnh trn, hnh trng hoc khng u.
T bo cht c c hn, chim gn ht hng cu.
Nhn chia : 16 32 mnh trng, sp xp khng u.
Ht sc t nu en th t li gia.
Khi th phn lit pht trin y , mi mnh trng c 1 vng TBC
bao quanh.
Th giao bo
Gametocyte

Hnh thoi, bu dc (non) hnh tri chui, qu thn (gi).
+ Giao bo c :
* Hnh qu thn, u trn.
* Nhn di dng ht n mu khng gii hn r.
* Ht sc t mu nu en th nm ri rc trn TBC mu tm c.
+ Giao bo ci:
* Hnh qu chui hay li lim.
* Nhn mu tp trung gia, chung quanh l cc ht sc t nu
en, TBC mu xanh.

c im Plasmodium vivax
Hng cu b k sinh Phnh to, nht mu, mo m.
Bnh thng c 1 KST, trong hng cu c th c n 2 KST.
S hin din ca cc th Thng thy tt c cc th.

T dng tr
( Th nhn )
Early Trophozoite

C hnh nhn.
TBC mnh mu xanh nht, c khng bo to, trn.
C 1 nhn nh bt mu .
T dng gi
Late Trophozoite

C dng amp
TBC nhn nheo c nhiu dng gi tc, khng bo b ct thnh
nhiu khng bo nh, c cc ht sc t vng nu to, nh khng
u.
Nhn to xp
C ht Schuffner mu hng phn b u trn mng hng cu.
Hng cu b k sinh to ln.
Th phn lit
Schizont

C hnh trn.
T bo cht chim ton b hng cu.
Nhn chia : 16 24 mnh trng, sp xp khng u.
Ht sc t nu en tp trung gia TBC.
Th giao bo
Gametocyte

Giao bo c :
Hnh cu, chim gn ht th tch hng cu.
TBC mu xanh xm
Nhn kh to, mu hng nht, thng gia.
Cc ht sc t vng nu xen ln ht Schuffner hng
trn TBC.
Giao bo ci:
Hnh cu
TBC xanh m.
Nhn nh gn c mu m, thng nm ra TBC.
Cc ht sc t vng nu xen ln ht Schuffner hng trn TBC.

c im Plasmodium malariae
Hng cu b k sinh Hi teo nh v sm mu.
Mt hng cu thng c 1 k sinh trng.
S hin din ca cc
th
Thy tt c cc th.
T dng tr
(Th nhn)
Early Trophozoite

C hnh nhn.
TBC dy mu xanh m, c khng bo to, trn.
C 1 nhn to mu thng lt vo khng bo bn trong.
TBC thng c ht sc t, mu nu en th.


T dng gi
Late Trophozoite

TBC b ko di hnh di bng vt ngang qua hng cu.
Ht sc t nu en th nm t li bn ra t bo cht.
Nhn to xp di thng nm trn ra TBC.
Hng cu b k sinh nh i.

Th phn lit
Schizont

C hnh trn
TBC chim ton b hng cu.
Nhn chia : 6 12 mnh trng, sp xp u n quanh khi sc
t nu en tp trung gia TBC to thnh th hoa hng, hoa cc.
Th giao bo
Gametocyte

Hnh trn hay bu dc, cu to ging Plasmodium vivax nhng kch
thc nh hn v hng cu b thu nh li.
Giao bo c nh hn giao bo ci.
Giao bo c: nh hn.
Hnh cu, chim gn ht th tch hng cu.
TBC mu xanh xm, bn trong ri rc cc ht sc t.
Nhn kh to, mu hng nht, thng gia.
Giao bo ci: hnh cu.
TBC xanh m.
Nhn nh, gn c mu m, thng nm ra TBC.

c im Plasmodium ovale
Hng cu b k sinh Hnh bu dc, nht mu, mo m.
Bnh thng c 1 KST.
S hin din ca cc th Thng thy tt c cc th.
T dng tr
( Th nhn )
Early Trophozoite
C hnh nhn.
T bo cht dy.
C 1 nhn to bt mu .


T dng gi
Late Trophozoite

C dng amp, hnh thoi khng u hoc b c rng ca.
T bo cht dy, c cc ht sc t vng nu
Nhn to xp.
C ht Schuffner mu hng .
Hng cu b k sinh phnh to ln.
Th phn lit
Schizont

Hnh bu dc.
TBC chim ton b hng cu.
Nhn chia: 8 12 mnh trng, sp xp u n quanh khi sc
t.
Ht sc t to mu nu en, tp trung gia TBC.
Th giao bo
Gametocyte

Ging P. vivax nhng nh hn.
Hnh cu, hnh bu dc.
C ht sc t th, vng nu xen ln ht Schuffner hng trn
TBC.



2.2. So snh hnh th ca cc loi Plasmodium theo tng giai on (hnh 30.6)
Th loi Plasmodium falcifarum Plasmodium vivax Plasmodium malariae

Hnh nhn, gn.

Nhn trn thm
hay tm.
Hnh nhn to.

Nhn to mu thm
hay tm.
Hnh nhn ging
P. vivax.
Nhn th v chc hn
P. vivax.
2.3. Cc c im xc nh cc loi Plasmodium trn tiu bn
Ngoi vic cn c vo cc c im chung v hnh thi, chng ta cn phi da vo mt s cc c im
ring bit ch yu ca tng loi Plasmodium d phn bit hn.

T dng
tr
TBC mnh, mu xanh da tri
hay xanh tm.
Chim 1/4 - 1/5 hng cu, c
th nm ra HC (th kt dnh
ngoi vi).
Mt hng cu c th c t
2 3 KST.
TBC dy v ln hn
P. falciparum.
Vng nhn dy ln
chim 1/3 2/3 hng
cu.

Trong hng cu c th
c n 2 KST (bnh
thng c 1 KST).
TBC thng c ht sc t,
mu nu en, th.

Mt hng cu thng c
1 KST.

T dng
gi
Hnh nhn, to.

TBC c, cc ht sc t
nu en, th, t li thnh cm.

C chng loi c ht Maurer,
mu nu, hng, c th gp
trong st rt c tnh.
Nhn to xp.

TBC c nhiu dng
gi tc

Trn TBC c cc ht
sc t vng nu, to nh
khng u nm ri rc
chen ln vi cc ht
Schuffner mu hng.
Nhn to xp, di thng
nm trn ra TBC.
TBC hnh trn, di bng
hay khn qung nm vt
ngang hng cu.
Ht sc t nu en, th
nm t li bn ra TBC.

Th phn
lit
C 16 32 mnh trng.
Cc mnh trng sp xp
khng u, cc ht sc t nu
en, th t li gia hay mt
bn m mnh trng c th
gp trong SRAT.
C 16 24 mnh trng.
Cc mnh trng sp
xp khng u xen k
vi cc
ht sc t vng nu
mnh
ri rc.
C 6 12 mnh trng
Cc mnh trng sp xp u
n bao quanh m sc t
nu en nh hoa th, hoa
hng.

Th giao
bo
Hnh thoi, bu dc (non)
hnh tri chui, qu thn (gi).

Giao bo c:
Hnh qu thn, ht u.
Nhn khng gii hn r.
Cc ht sc t mu nu en,
th nm ri rc trn TBC.
Giao bo ci:
Hnh qu chui hay li
lim.
TBC xanh nht bao quanh
nhn tm.
Cc ht sc t en, th ri
rc trn TBC.
Hnh trn, bu dc
TBC to gn chim
gn ht hng cu.

Giao bo c:
Nhn to, xp, gii
hn khng r nm
gia TBC.
Cc ht sc t vng
nu nm ri rc trn
TBC xen ln cc ht
Schuffner hng .
Giao bo ci:
Kch thc ln
Nhn gn nh, chc,
gii, hn r nm ra
TBC.
Cc ht sc t vng
nu xen ln ht
Schuffner hng trn
TBC.
Hnh trn hay bu dc.
Cu to ging Pl. vivax
nhng kch thc nh hn.
Cc ht sc t th, m
mu.

Giao bo ci ln hn
giao bo c.

c im P. falcifarum P. vivax P. malariae

3. NHNG BIN I V HNH TH K SINH TRNG ST RT
3.1. Nguyn nhn
Do thuc iu tr, thuc nhum, cht khng ng hoc do k thut lm tiu bn v nhum.
3.2. S bin i v hnh th
Nhn c th c v bt mu m hn, hay tr nn xp hn v bt mu nht hn bnh thng.
Nhn c th to ln hoc teo nh li, hay v thnh tng mnh vn.
TBC bt mu nht hay hng (thay v xanh), c th phnh to, teo nh hay pht trin cc dng bt
thng.
Kch thc KST thu nh hay phnh to ra.
Ht sc t, ht c hiu i khi khng xut hin.

CU HI LNG GI
1. Anh (ch) cho bit nhng thnh phn cu to c bn ca mt KST SR?
2. Anh (ch) c nhn xt g v hng cu nhim Plasmodium falciparum v Plasmodium vivax?
3. Anh (ch) da vo yu t no xc nh bnh nhn nhim P. falciparum hay P. vivax khi quan st tiu
bn mu nhum?
4. C khi no th phn lit ca P. falciparum xut hin trong mu ngoi bin? Anh (ch) cho bit trng hp
no? Trong trng hp ny, anh (ch) lm g bit c bnh nhn ny nhim P. falciparum hay P.
vivax?
5. Anh (ch) nu im khc bit phn bit th t dng gi v th giao bo ca Plasmodium vivax.
6. Th t dng gi ca P. malariae c c im no khc vi nhng loi KST SR khc?
7. Nhn dng KST SR trn git mu mng d hn trn git dy, nhng phng xt nghim, cc k thut
vin thng thch quan st git dy hn git mng. Anh (ch) gii thch ti sao?

Hng cu
b k sinh
Tng i bnh thng. Phnh to, nht mu mo
m.
Hi teo nh v sm
mu.
S hin din
ca cc th
Thng thy th nhn v
th giao bo (th t dng
gi v phn lit c th gp
trong SRAT).
Thng thy tt c cc
th.
Thng thy tt c
cc th.
Ht sc t Hnh que, th, en, nu en,
thng t li.
Hnh que, mnh vng nu,
nh vng nm ri rc u.
Hnh que th, nu en,
tp trung hai bn ra
TBC.
Ht c hiu Ht Maurer nu, ln,
tha tht (ch c mt s
chng loi).
Ht Schufffner mu hng,
nh v ri u (thng
gp).
Ht James hay
ht Ziemann (nhum
c bit mi thy).

Bi 31
HNH TH TIT TC Y HC

1. I CNG
Tit tc l nhng sinh vt a bo, khng xng sng; c th i xng, phn t, c bao bc bi v
cng kitin. Chn gm nhiu t c ni vi nhau bng khp.
Ngnh Tit tc chim khong 80% cc loi ng vt trn qu a cu ny. Chng trc tip gy bnh
hoc truyn bnh cho ngi v ng vt. Giai on trng thnh v u trng u c th gy bnh cho ngi
bng cch chm nc c, gy d ng nga, ht mu, xm nhp m cng nh truyn cc bnh vi trng, virus
v k sinh trng.
2. C IM CHUNG V CU TO CA TIT TC
2.1. S phn t
C th phn t. Lp cn trng c s phn t r rt, lp nhn s phn t khng r.
2.2 V kitin
L b xng ngoi, gip tit tc chng cc tc ng ca ngoi cnh, lm im ta cho h c v c quan
chuyn ng hot ng linh hot. Lp v kitin bao ph c th khng ng u tng phn, thng dy hn
phn ngc, nhng ch dy ln gi l mai, hoc khin, hoc gip.
V kitin cng cn tr s pht trin ca tit tc: tit tc ch ln ln c khi v kitin cn mm, khi v
kitin tr nn cng, n khng ln c na, lc phi thay v c, l hin tng lt xc. V vy, trong
qu trnh pht trin, tit tc phi c giai on lt xc thay i hnh dng v gia tng kch thc.
2.3. Cu to c th tit tc
2.3.1. Cu to bn ngoi
C th tit tc gm c 3 phn: u, ngc, bng.
Phn u c th pht trin hon chnh, gm mt, ru, xc tu, ming, vi, Mt s tit tc c u
khng hon chnh gi l u gi.
Cc gic quan:
+ Mt: c th l mt n hoc mt kp. Tit tc thuc lp nhn khng c mt hoc nu c l mt n.
Tit tc thuc lp cn trng bao gi cng c mt kp.
+ Xc bin hm, c nhim v gi thng bng cho c th, tm vt ch, tm ch ht mu.
+ Ru, c nhim v nh hng.
Phn ngc mang cc c quan vn ng nh chn, cnh.
Phn bng chia lm nhiu t, nhng t cui to thnh b phn sinh dc ngoi.
Ton thn tit tc cn c th c vy hoc lng to thnh nhng khoang mu khc nhau.
2.3.2. Cu trc bn trong
Bn trong c th l xoang thn, cha cc c quan ni tng.
H tiu ha: thng pht trin hon chnh, gm ming, thc qun, rut, hu mn, tuyn tit cht
ng. Phn ph ming c nhiu bin i thch nghi vi nhng thc n khc nhau.
H tun hon gm c tim, c hnh ng di vi nhng on phnh thnh ti v l tim mu tr v tim.
H mch h, mu t tim chy vo xoang gia cc c quan.
H thn kinh gm nhng dy thn kinh, hch thn kinh, c th c hch thn kinh trung tm lm nhim
v no.
H bi tit l nhng th hnh ng sp xp mt s t nht nh. Lp nhn v cn trng c nhng ng
Malpighi lm nhim v bi tit.
H sinh dc: b phn ngoi pht trin n mc hon chnh.
+ B phn sinh dc ci gm c 2 bung trng, ng dn trng, m o v ti cha tinh, gip cho s th
tinh c nhiu ln, mt s tit tc c c nt giao hp lm nhim v bo qun tinh trng.
+ B phn sinh dc c c 2 tinh hon, ti tinh, tuyn ph, ng phng tinh v gai giao hp.


3. PHN LOI TIT TC
Cc tit tc c chia lm nhiu lp, nhng ch c 3 lp c lin quan n y hc. Di y l bng phn
loi tm tt.

Bng so snh cc c im c th v sinh hc
ca 3 lp cn trng, nhn v gip xc
c im
Insecta (Cn trng) Arachnida (Nhn) Crustacea (Gip xc)
Ch, rn, rp, b cht, rui
mui.
Ve, m, mt, ci gh, b cp. Tm, cua, thy tao (Cyclops).
C th
Chia lm 3 phn: u,
ngc v bng.
Chia lm 2 phn: u ngc
v bng.
Phn t a dng, thng c
2 phn: u ngc v bng.
u 1 i ru. Khng c ru. 2 i ru.
Chn 3 i chn.
Trng thnh c 4 i chn, S lng chn thay i,

LP NHN
(Arachnoidea)

Lp ny c nhiu b khc nhau nhng ta ch hc b Ve m (Acarina)
A. C IM CA LP NHN
C th chia 2 phn: u ngc v bng.
C 8 chn lc trng thnh, u trng ch c 6 chn.
Khng c cnh, khng c ru.
C b phn ming bin i c th chch c gi l u gi gm c:
+ 1 h khu pha bng hnh qu da c nhiu gai hng v sau.
+ 2 cng pha lng hnh kp hay mc.
+ 2 xc bin hnh chy ng vai tr che ch nm hai bn.
B. PHN LOI
B Ve m chia ra lm 2 nhm:
Nhm ht mu gm c cc h Ixodidae, Argasidae v Trombiculidae.
Nhm sng da, ch yu c h Sarcoptidae.

1. H Ixodidae thng gi l ve cng, c nhng c im sau y:
Ve trng thnh, c th l mt khi: u, ngc, bng khng phn chia.
B phn ming gi l u gi.
u gi c 1 phn nh ra hnh qu da, c nhiu gai mc ngc, c 2 cng khng di ng v 2 xc
bin.
Trn thn ve c nhng vng lp kitin dy ln thnh tng tng gi l khin (mai). Ve ci c khin lng
nh, khng c khin bng. Ve c c khin bng v khin lng rng che ht lng.
Mt bng ve c l sinh dc pha trc v hu mn pha di.
L th hai bn gc chn th t.
Chn gm nhiu t, ngn chn cui cng c mng v c gai bm. i chn th nht c b phn c
chc nng nh c quan khu gic.
Trong h ny ta hc cc ging sau y: Ixodes, Rhipicephalus, Dermacentor m ta phn bit nh
nhng c im sau y:
Rnh bc hu mn pha trc hu mn, u gi di: Ixodes.
Rnh bc hu mn pha sau hu mn, chy ngn, chn u gi hnh
6 gc: Rhipicephalus.
Rnh bc hu mn pha sau hu mn, chy ngn, chn u gi hnh ch nht, hu mn g ln, chn
u trng c 3 i chn. thng c 5 i chn.
Bng C nhiu t.
Cnh C hoc khng cnh. Khng c cnh. Khng c cnh.
Th Bng ng kh. Bng ng kh hoc phi Bng mang.
Vai tr trong
y hc
Gy bnh hoc trung gian
truyn bnh.
Gy bnh hoc trung gian
truyn bnh.
K ch trung gian ca nhiu
loi giun, sn ca ngi v
ng vt.
chy khng gc cnh: Dermacentor.

2. H Argasidae cn gi l ve mm, c nhng c im sau y:
Khng c mai lng.
u gi nm mt bng, khng nh ra ngoi.
t chn cui cng ch c 2 mng.
Trong h ny c 2 ging chnh:
+ Argas, thn dp, c mt ln phn chia r rt gia mt lng v mt bng.
+ Ornithodorus, thn to v dy, khng c ln phn chia r rt gia mt lng v mt bng.

3. H M (H Trombiculidae)
H Trombiculidae c l th pha trc thn.
Ch u trng Trombicula ht mu v c th truyn bnh.
u trng rt nh ( 200m) c mu hoc mu vng, thn c nhiu lng t.
C u gi.
C khin lng.
C 3 cp chn, t cui c 3 mng.
Con trng thnh: 1m, c hnh dng tng t nh chic n ghi ta vi
8 chn, thn c nhiu lng hn dng u trng, sng t do v ht nha cy.

4. Mt (Dermanyssus gallinae)
Thng k sinh g, gi l mt g.
Thn c hnh qu l, mu trng hoc mu ty theo s thay i ca mu trong thn.
u gi c hnh kim, xc bin c khp c ng c.
C th truyn bnh toi g v bnh vim mng no cho ngi v nga.

5. H Sarcoptidae
Ph bin v k sinh c Sarcoptes scabiei gy bnh gh nga.
Ci gh trng thnh:
+ Thn hnh bu dc, lng g.
+ Kch thc: con ci: 300 400m, con c 200250m.
+ Mu vng nht.
+ Ming rt ngn, khng c mt.
+ Khng c l th v th qua da mng.
+ C 4 cp chn:
* Con ci: cp chn th 4 tn cng bng lng t.
* Con c: cp chn th 4 tn cng bng mt ng ht.
u trng: ging con trng thnh, nhng c 3 cp chn.

LP CN TRNG

A. C IM CA LP CN TRNG
Cn trng l nhng tit tc c ru.
Sng trn mt t hay trn khng.
C 3 cp chn.
Thn chia lm 3 phn r rt: u, ngc v bng.
u c mt, ru v cc b phn ming (1 mi trn, 2 hm trn, 2 hm di v 1 mi di v c thm
xc bin).
Ngoi ra, mt s loi cn trng, cn c nhng chi bng gic t xut pht t yt hu:
thng yt hu dnh vo mi trn, h yt hu cha ng ng nc bt.
Tt c cn trng k sinh u c b phn ming kiu chch hay ht.
Ngc c 3 t, mi t c mt cp chn. Hai t cui c th c mt hay hai cp cnh.
Bng thng rt n nang v trung bnh c t 8 n 9 t.

B. PHN LOI LP CN TRNG
Nhng cn trng k sinh ngi thuc 4 b sau y:
B Diptera: c 2 cnh th mng (rui, mui).
B Anoplura: khng c cnh.
B Hemiptera: c 4 cnh.
B Siphonaptera: khng c cnh.
B Diptera v b Siphonaptera c chu k bin thi hon ton (hnh th ca u trng khc hn con
trng thnh).
B Hemiptera v b Anoplura c chu k bin thi khng hon ton (hnh th ca u trng gn ging con
trng thnh).
1. B Diptera chia lm 2 b ph da vo s t ca ru:
B ph Tim gic (Nematocera): ru di, c trn 3 t.
B ph on gic (Brachycera): ru ngn, c di 3 t.
1.1. B ph Tim gic (Nematocera)
a) Mui trng thnh
Mui c tm quan trng nht v phng din y hc v c nhng c im sau y:
+ u mui c hnh cu, mang 2 mt kp, vi, xc bin v ru.
+ Vi kiu chch, gm c: mi di v mi trn rt n nang un cong li to thnh mt ci vi, cn hm
trn v hm di bin thnh nhng trm bn nhn c th xuyn thng da.
+ Xc bin: hai bn ca vi, c chc nng xc gic. Xc bin khc nhau ty theo ging v loi mui
nn dng nh loi.
+ Ru ca con c v con ci khc nhau: ru ca con c c nhiu lng v rm, ru ca con ci tha v
ngn.
+ Ngc mui gm 3 t, mi t mang mt i chn, t gia mang thm i cnh.
+ Cnh mui c cc ng sng dc v ng sng chch costa. Trn cnh c vy, vy to nn rim
cnh. Nhng ng sng trn cnh mui v hnh th vy trn cc ng sng c gi tr trong nh loi mui.
+ Bng c 9 t, t cui l b phn sinh dc. Gia cc t bng c cc bng mu do vy to nn.
+ Chn rt di v mnh, gm nhiu t: t hng, i, cng v bn. Bn chn c 5 t, tn cng bng
vut.

MUI CI MT LNG

C 4 ging chnh: Anopheles, Culex, Aedes v Mansonia.
Phn bit 4 ging ny da vo c im ca mui trng thnh, u trng v trng.

chy

b) u trng (B gy)
u trng khng c chn, di, c nhng chm lng ta ngang, xp t i xng dc theo thn.
u trng c 4 giai on, hnh th ging nhau, ch khc nhau v kch thc, u trng giai on 4 c
chiu di 1cm.
u hi dt, c mt kp, ru rm, ming nhai c.
Ngc, bng cng c nhiu lng. Hai t bng cui bit ha thnh b phn th: 2 l th Anopheles;
ng th Culex, Aedes v Mansonia.


c) Trng
Hnh th ca trng ty tng loi mui.
Trng ca Anopheles: hnh bu dc, ri rc, c phao 2 bn.
Trng ca Aedes: hnh thoi, ri rc.
Trng ca Culex: hnh thoi, kt thnh b.
Trng ca Mansonia: hnh thoi, c gai 1 u, dnh thnh chm.

C IM PHN BIT
Anopheles spp Aedes spp Culex spp Mansonia spp


c im Anopheles Culex Aedes Mansonia
Mui
trng
thnh
Xc bin
hm
Di bng vi,
phnh to cui
con c,
thun con ci.
Di hn vi
con c, ngn
hn vi con
ci.
Ging nh
Culex, nhng
gia cc t
chn c nhng
khoang trng.
Ging nh
Culex.

Cnh Thng c m Khng c m. Khng c
m.
Khng c m,
c vy rng.
Mu sc Mu nu, vng
nu hoc nu
sm.
Ngc lng,
chn v thn c
nhng m
hoc khoanh
mu trng, en.
C mu vng
rm, ngc, chn
v thn c
nhng m
trng.
T th u Chch (xin) vi
im ta.
Song song vi
im ta.
Song song vi
im ta.
Song song vi
im ta.
t cht ca
bng
Trn. Trn. Nhn.
u trng B phn h
hp t
th 8 ca
bng
C 2 l th. ng th di v
hp.
ng th ngn
v rng.
ng th tn
cng bng gai
nhn, cm vo
r cy.
T th di
nc
Nm ngang vi
mt nc.
Nm chch vi
mt nc.
Nm chch vi
mt nc.
Cm vo r cy
thy sinh.
Trng Hnh thoi, ring
r v c phao
hai bn hng.
Hnh tr, mt
u hi nhn
v kt thnh b
ni trn mt
nc.
Hnh thoi, ring
r khng c
phao.
Hnh thoi, c gai
mt u, dnh
thnh chm v
cm vo mt
di l cy.

1.2. B ph on gic (Brachycera)
Bao gm cc loi rui chch ht mu v khng ht mu.
Ru ngn, 2 cnh rng, chn ngn v n nang.
a) Rui nh (Musca domestica)
Con trng thnh:
+ Di t 6 7m, c thn mu en xm, bng mu vng xn.
+ u hnh bn cu c 2 mt kp, cp mt con c gip lin nhau, cn cp mt con ci cch xa nhau.
+ Pha trc u c ru ngn 3 t, c xc bin hm v vi.
+ Vi thuc kiu lim, u vi xo ra, ni y c 2 phin cha nhiu ng ht nh c th ht cc dch
hu c.
+ Ngc gm 3 t, mang 2 cnh v 6 chn, cnh rui trong sut.
+ Bng rui c gm 8 t, bng rui ci c t cui tr thnh b phn sinh dc. B phn sinh dc
ca rui ci thng b co n vo pha trong, khi mi mi thy r.
u trng rui (gii):
+ Mu trng ng, khng c lng, khng c chn, u nhn, ui bu, thn c vn, gm nhiu t.
+ Ming c 2 mc, l th pha trc c hnh ngn tay, l th pha ui c hnh dng khc nhau ty
theo ging.

b) Rui tru (Tabanus sp)
Rui c thn hnh ln, kch thc 1,5 2cm.
Ton thn c ph bng lng mn, ngc en c vch vng, bng mu vng.
u to, mt to, b phn ming kiu chch, ru d thy. Con c c 2 mt st nhau, con ci 2 mt
cch xa nhau.
Vi rng, chc xung di.
Cnh trong sut, hoc c mu hi nu, c vn. Khi ngh, cnh cp xung st thn.

2. B Aphnaptera
2.1. c im chung
C thn dp theo chiu dc, kch thc t 1 5m, cnh teo li v hai chn sau n nang.
C ming kiu chch.
B cht trng thnh mu vng, kch thc 1 6m, u hnh bu dc, u dnh lin vi ngc.
u c mt n, ru, xc bin mi c 4 t.
Phn di u ca mt s ging c nhng lng nhn, cng, to v xp thnh hnh rng lc, nn c
gi l lc.
Ngc gm 3 t, mi t mang 1 i chn. i chn th 3 to, khe v di dng nhy. Lng t
ngc 1 i khi c mang lc.
Bng gm 10 t. t 8, 9 dnh vi nhau v mang b phn sinh dc. Con c c rnh sinh dc t
cui, con ci c ti cha tinh hnh mc o pha ui. t th 10 cha hu mn v 1 c quan cm gic.

2.2. Phn loi b cht
Phn loi b cht da vo lng v lc:
a) B cht khng lc
Pulex irritans: 1 lng trc mt pha di mt v 1 lng sau u.
Xenopsylla cheopis: lng trc mt ngang vi mt, lng sau u nhiu v xp thnh hnh ch V.
b) B cht c 1 lc
Nosopsyllus fasciatus c mt lc ngc (lc gy).

c) B cht c 2 lc
Ctenocephalides canis v Ctenocephalides felix c hai lc, mt u, gn ming (lc m) v mt
ngc (lc gy).


3. B Anoplura
Gm nhng cn trng di t 2 3m, khng c cnh, chn n nang, tn cng bng nhng mng cong to
bm vo da, thn dp theo chiu ngang, c b phn ming kiu chch.
3.1. Ch (chy) (Pediculus humanus)
a) Con trng thnh
Sng k sinh ngi, thng tc (var capitis), hay bm vo qun o (var corporis).

Thn di, dt theo chiu lng bng, di 2 4mm.
C mu xm hoc nu.
u tch ring vi phn ngc v gm 2 mt n, 2 ru c 5 t.
Vi ngn khng trng thy v tht vo trong u.
Ngc c 3 t nhng khng phn bit r rng, gia ngc c 2 l th, ngc mang 6 chn.
Bng c 9 t, t t 1 n t 6, mi t c 1 cp l th 2 bn thn. Nhng t cui ca bng c
b phn sinh dc:
+ Con c thng cui bng hi nhn, c mt gai to nh ra ngoi
+ Con ci cui bng c 2 thy, c mt l sinh dc.
b) Trng
Trng hnh bu dc, di 0,8m, c np vi 1 hng t bo.

3.2. Rn (Phthirus pubis)
Con trng thnh di 1,5 2m. Thn ngn, ngc rt rng, b ngang ca ngc to hn bng v ngc
bng khng phn chia r rt. Bng ngn v ch c 5 t.
u tng i ngn, nm tht trong lm ca ngc.
Chn to mp, c mng di, to cong li u nn bm rt chc vo ni
k sinh.

4. B Hemiptera
Gm nhng cn trng 4 cnh nhng cnh c th b thoi ha v mt i. Trong b ny, ta ch hc rp
(Cimex lectularius): ht mu, thng sng cc khe g v rp c cnh (Triatoma sp).
4.1. Rp (Cimex lectularius)
Rp mu nu sm, di 3 6mm.
Thn mp b ngang, lng v bng dt, khng c cnh.
u nh, dt, hnh nm gc, tht vo trong mt lm ca ngc v c 2 mt li v mang 2 ru c 5 t.
Ngc gm 3 t, mi t mang 1 i chn. Ngc gia c mt cp cnh rt th s, c hnh bu dc.
Bng hnh bu dc, c 11 t, ch thy r 8 t, nhng t cui cng thnh b phn sinh dc ngoi.

4.2. Rp c cnh (Triatoma sp)
C kch thc to.
Rp c mu nu m, nh vng ngc, cnh v 2 bn bng.
u di v hp, c mt i mt kp li ln, 2 mt n, ru c 4 t, vi mnh, gp v pha bng.
Thn hp, dp, c mang cnh. Chn di c 3 t.

CU HI LNG GI
1. Anh (ch) m t hnh th chung ca tit tc.
2. Nu nhng im khc bit gia tit tc lp cn trng v lp nhn.
3. Anh (ch) nhn dng ve da vo nhng c im no? Ve cng khc ve mm im no?
4. M t hnh th ca ci gh.
5. M t hnh th ca mui.
6. Nu cc im ging v khc nhau ca mui Anopheles, Culex, Aedes v Mansonia.
7. Phn bit u trng (b gy) ca Anopheles, Culex, Aedes v Mansonia.
8. So snh trng ca Anopheles v Aedes, trng ca Culex v Mansonia: ging v khc nhau im no?
9. M t cu to c bn ca loi rui. Rui khc mui im no?
10. M t b cht trng thnh.
11. Phn bit b cht chut, b cht ngi v b cht ch, mo.
12. Ch khc vi rn im no?


Bi 32
CHN ON VI NM NGOI DA

1. MM BNH: Nm si
1.1. Microsporum c khong 15 loi:
Microsporum canis, Microsporum audouinii, Microsporum gypseum, Microsporum ferrigineu,
1.2. Epidermophyton ch c mt loi: Epidermophyton floccosum.
1.3. Trichop hyton c khong 21 loi: Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton
concentrium, Trichophyton violaceum, Trichophyton tonsurans, Trichophyton verrucosum,
2. BNH PHM
Da: co da phn ngoi vi sang thng.
Mng: co phn li xi nh bng ci di mng.
Tc: chn nhng si tc xp, mt mu, d gy hoc nhng si tc pht hunh quang di nh sng
n Wood.
3. KHO ST TRC TIP
Bnh phm dm vo KOH 20%, kho st di knh hin vi.
3.1. Mu da, mng: trn nn t bo biu b ca da v mng c nhng si t nm phn nhnh, phn vch,
i khi si t nm t thnh tng on ngn.

3.2. Mu tc: c th gp 5 kiu tc nhim nm v c chia lm hai nhm chnh:
a) Nhm pht ni ngoi
Kiu Microsporum: bao quanh si tc l mt lp nhiu bo t trn ng knh khong 2mm, xp u
n, bn trong si tc l si t nm.
Kiu ging Microsporum: bao quanh si tc l cc bo t ng knh (d) bng 3 4m, xp khng
u, i khi ri rc thnh chui, bn trong si tc c si t nm.
Kiu Microsporum bo t ln: ging nh kiu Microsporum nhng bo t ln d = 5 6m.

b) Nhm pht ni
Kiu pht ni: bn trong si tc cha y cc bo t trn, d = 3 4m.
Kiu lm chn: bn trong si tc c cc si t nm c th t thnh on ngn, khi h nng bn trong
si tc c nhiu bt kh.

3. CY NM
Bnh phm c cy ln mi trng Sabouraud c chloramphenicol v Cycloheximide (mi bnh nhn
cy 3 4 ng). Mt thi gian sau vi nm s mc thnh khun lc nm (7 10 ngy 1 thng 1,5 thng).
Da vo c im, mu sc, tc mc v hnh th ca nm cng cc xt nghim khc nh danh nm.
Cy ln mi trng cm ngui phn bit Microsporum canis vi Microsporum gypseum
(Microsporum canis mc mnh).
Cy ln mi trng ur; mi trng thch khoai ng v th nghim xuyn tc phn bit hai loi
vi nm Trichophyton rubrum v Trichophyton mentagrophytes.
+ Mi trng ur: trong vng 7 ngy.
Trichophyton mentagrophytes: (+) mi trng i mu .
Trichophyton rubrum: () mi trng khng i mu.
+ Thch khoai ng:
Trichophyton mentagrophytes: khng sinh sc t.
Trichophyton rubrum: sinh sc t nho.
+ Th nghim xuyn tc: quan st la cy sau 10 ngy
Trichophyton mentagrophytes: xuyn tc (+): vi nm n thng si tc.
Trichophyton rubrum: xuyn tc ().

Cc loi Trichophyton: cy ln mi trng Trichophyton 1 7 ngy nh danh tng loi nm.
c im khun lc nm v hnh th quan st di knh hin vi ca mt s vi nm thng gp:
+ Trichophyton rubrum:
* Khun lc nm nhuyn nh bt, c sc t nho.
* Hnh th quan st di knh hin vi: si t nm mnh, hai bn c cc bo t nh nh hnh git nc,
i khi c bo t nh ln.

+ Trichophyton mentagrophytes:
* Khun lc nm nhuyn nh bt hay mt nh nhung, c sc t mu kem, i khi .
* Hnh th quan st di knh hin vi: si t nm mnh, i khi c nhng si xon, dc hai bn nhnh
nm c bo t nh nh, trn, i khi c bo t nh ln vch nhn.


+ Trichophyton schoenleinii:
* Khun lc nm nh sp, g cao v c nhiu np nhn.
* Hnh th quan st di knh hin vi: ch thy si t nm c vch ngn, khng thy bo t.
+ Microsporum audouinii:
* Khun lc nm mt nh nhung, sc t mu hng nht.
* Si t nm cong queo, nhiu bo t bao dy, c th c si t nm hnh vt, him khi c bo t nh
ln.
+ Microsporum canis:
* Khun lc nm mt nh nhung, sc t cam, mc rt tt trn mi trng cm.
* Hnh th quan st di knh hin vi: nhiu bo t nh ln hnh thoi, vch dy, c gai, bo t c 5
8 vch ngn.
+ Microsporum gypseum:
* Khun lc nm nhuyn nh bt, sc t vng, nu nht.
* Hnh th quan st di knh hin vi: nhiu bo t nh ln hnh thoi, vch mng, c
gai, 5 8 vch ngn, ngoi ra cn c bo t nh nh hnh qu l.

+ Epidermophyton floccosum:
* Khun lc nm nhuyn nh bt, sc t vng xanh l cy nht.
* Nhiu bo t nh ln hnh chy, 3 4 vch ngn, thng dnh chm
2 3 ci.

CU HI LNG GI
1. M t hnh nh hin vi ca vi nm trong mu da.
2. M t hnh nh hin vi ca vi nm trong mu tc.
3. Nu c im phn bit vi nm gy bnh lang ben v vi nm ngoi da trong mu da.
4. Nu c im phn bit vi nm gy bnh trng tc en v vi nm ngoi da trong mu tc.
5. M t hin tng pht ni v pht ngoi.
6. Chn on phn bit vi nm: Trichophyton rubrum v Trichophyton mentagrophytes.

Bi 33
CHN ON VI NM NGOI BIN

1. LANG BEN (Tinea versicolor)
Tn mm bnh: Pityrosporum orbiculare
a) Bnh phm
Chiu sng n Wood, vng da c vi nm s pht hunh quang mu vng nht.
Co vy da cho rt vo hp Petri, trn lam knh, hoc giy trng kh, sch.
C th dng bng keo trong dn ln vng da lang ben dng bong vy.
b) Kho st trc tip
Vy da dm vo trong KOH 20%.
Bng keo trong dn ln lam knh.
Lm pht t vi dung dch KOH 20% v quan st di knh hin vi, ta thy nm hin ra thnh tng
m nh gm c si t nm v t bo ht men. Si t nm ngn v cong, ng knh khong 3m, phn
nhnh thnh hnh ch S, V hay Y; t bo ht men trn ng knh 3 8m xp thnh m 10 30 t bo.

2. TRNG TC EN (Black piedra)
Tn mm bnh: Piedraia hortai.
a) Bnh phm
Nh nhng si tc c nhng ht en nh cng bm cht ln si tc
b) Kho st trc tip
Lm pht t: ct tc thnh on ngn 1 1,5cm, dm vo KOH 20% em kho st di knh hin vi
thy nhng ht nh c cu to bi nhng si t nm mu nu ng knh bng 4 8 m, vch dy, vch
ngn gn nhau. Cc si t nm an kt vi nhau thnh 1 bao, bn trong l nhng ti trn, ng knh 30
60m, bn trong ti trn cha 2 8 bo t ti hnh qu chui, di 13 31m, mi u c 1 si dy nh (roi).

CU HI LNG GI
1. Vit tn nm lang ben v trng tc en.
2. M t hnh nh di knh hin vi ca nm lang ben mu da.
3. M t hnh nh di knh hin vi ca trng tc en trn mu tc.

Bi 34
CHN ON VI NM CANDIDA

1. MM BNH
C 8 n 9 loi Candida c kh nng gy bnh nhng trn 95% trng hp l do Candida albicans.
2. BNH PHM
Rt a dng, ty theo dng bnh m ta c nhng bnh phm khc nhau: mng trng trong ming, m,
da, mng, dch m o, nc tiu, phn, mu, sinh thit cc c quan.
Bnh phm ly xong phi gi n phng xt nghim ngay trong vng 24 gi.
3. KHO ST TRC TIP
Dch m o, mng trng trong ming, m, cn lng nc tiu, kho st vi NaCl 0,85% hoc pht
ln lam nhum Gram.


Da, mng, sinh thit kho st vi KOH 20%.
Cc mu sinh thit: sau khi ly c b vo dung dch c nh, x l theo phng php m hc ri
em nhum Hematoxyline v Eosin (HE) hoc nhum Periodic Acid Schiff (PAS).
Khi kho st cc loi bnh phm di knh hin vi thy nhiu t bo ht men v si t nm gi.
4. CY NM
Cc loi bnh phm cy vo mi trng Sabouraud + Chloramphecol 25
O
C.

Mu mu: cy vo mi trng BHI 37
o
C trong 10 ngy. Sau cy chuyn sang Sabouraud +
Chloramphenicol. Khun lc nm Candida sp c mu trng nho.
4.1. nh danh Candida albicans
a) Th nghim sinh ng mm (Serum test)
Trong mt ng nghim c cha sn 0,5 1ml huyt thanh ngi hay th, ha tan mt t
khun lc nm hoc bnh phm vo huyt thanh, 37
o
C trong 4 gi. Ly mt git huyt
thanh nh ln lam knh, kho st di knh hin vi.
Nu thy c ng mm: Candida albicans.
Nu thy khng c ng mm: l cc loi Candida khc.

b) Th nghim tm bo t bao dy (Chlamydospore)
Cy vi nm ln thch bt bp, thch khoai ty c rt mt theo k thut Dalmau.
Quy trnh k thut:
+ Dng kim cy ly nm t mi trng Sabouraud.
+ Vch su vo trong thch hai ng song song, di 1,5cm.
+ t kim cy ri vch ng ziczac ln mt thch.
+ y l knh ln ng cy.
+ nhit phng 48 96 gi, kho st knh hin vi.
Kt qu:
+ Nu thy t bo men, si t nm gi, bo t bao dy: Candida albicans.
+ Nu thy t bo men v si t nm gi: l Candida sp khc.
+ Nu thy t bo men: khng phi Candida sp m l mt loi nm men khc.

4.2. nh tn cc loi Candida sp khc
a) Cy ln mi trng Sabouraud lng
Ha tan mt t nm vo ng Sabouraud lng.
nhit phng t 13 ngy.
Nu thy nm mc thnh lp vng trn mt mi trng, l Candida krusei hoc Candida tropicalis.
Nu nm mc thnh cn y ng nghim, l cc loi Candida khc.
b) Th nghim ln men ng
Dng 4 ng nghim c pepton lng, cht ch th mu tm Bromocresol purple (BCP), ng Durham.
Cho vo mi ng mt loi ng: Glucose, maltose, saccharose, lactose.
Cy nm vo 4 ng nghim ni trn.
thm hn hp du parafinvaselin ln trn to iu kin k kh.
37
0
C trong 10 ngy.

Khi ln men, nm s bin ng thnh acid v hi. Acid lm mu tm ca BCP chuyn sang mu
vng. Hi nu c sinh ra s bay ln lm h mc nc trong ng Durham.
c) Th nghim ng ha ng
Cy nm vo mi trng ng ha.
Khi mi trng ng th t ln mt thch cc a giy c tm ng glucose, maltose, saccharose,
lactose, galactose, raffinose.
nhit phng trong 2 ngy.
ng ha loi ng no, nm s mc quanh a ng .
d) Th nghim ng ha Nitrate
Quy trnh cng ging nh ng ha ng. Ch khc l khi mi trng ng th t ln mt thch a
KNO
3
.
Bng tm tt cc c im nh danh mt s loi Candida sp. thng gp

5. NGHA VIC TM THY CANDIDA TRONG BNH PHM
Nu bnh phm c tnh tp nhim nh nim mc ming, dch m o, m, phn: ch c ngha bnh
khi kho st trc tip thy nhiu si t nm gi v t bo men. i vi cc loi bnh phm ny kho st trc
tip quan trng hn cy.
Nu bnh phm l mu, dch no ty, ty xng, m t p xe, kho st trc tip hay cy tm thy
Candida u c ngha chn on.
Candida sp trong bnh phm.


CU HI LNG GI
1. M t hnh nh hin vi ca vi nm Candida sp. trong bnh phm khi soi ti, nhum Gram.
2. M t k thut tm bo t bao dy.
3. Trnh by quy trnh nh danh vi nm Candida albicans.
4. Nu ngha ca vic tm thy vi nm Candida sp. trong mu m, dch no ty.

Bi 35
CHN ON VI NM ASPERGILLUS

1. MM BNH
Do cc loi vi nm Aspergillus sp.
V d: A. fumigatus; A. nidulans; A. flavus; A. niger; A. terreus.
2. BNH PHM
Ty th bnh ta c nhiu dng bnh phm: m, gic mc, da tai, m tai, sinh thit phi hoc sinh thit
cc c quan,
3. KHO ST TRC TIP
m: nhum Gram, nhum khng acid, soi ti vi KOH 20%.
M: soi ti vi NaCl 0,85%, nhum Gram.
Gic mc: soi ti vi NaCl 0,85% (v trng).
Da tai: soi ti vi KOH 20%.
Sinh thit cc c quan: x l theo phng php m hc nhum Hematoxyline v Eosin (HE), Perodic
Acid Schiff (PAS) hoc Gomori.

Trn tt c cc loi bnh phm thy c nhiu si t nm d = 4 5m, phn nhnh, phn vch, khi phn
nhnh, nhnh ph to vi nhnh chnh gc 45
O
, i khi thy u bo i, cc tiu bo i v bo t.

4. CY NM
Bnh phm c cy trong mi trng Sabouraud c hay khng c Chloramphenicol 37
O
C v nhit
phng th nghim. Sau 3 4 ngy vi nm s mc thnh khun lc, cy chuyn sang mi trng c bit
cy vi nm Aspergillus l mi trng Czapek. Khun lc nm s pht trin v mi loi s c c im
khc nhau da vo nh danh.
Trong chn on bnh vi nm Aspergillus nhit cy quan trng phn bit vi nm Aspergillus gy
bnh vi Aspergillus hoi sinh.

Nhit
Tc mc
Aspergillus hoi sinh Aspergillus gy bnh
37
o
C

Nhit phng
Chm
Nhanh
Nhanh
Chm

CU HI LNG GI
1. M t hnh nh vi nm Aspergillus sp. trong mu m v gic mc di knh hin vi.
2. M t hnh nh ca vi nm Aspergillus sp. trong mi trng cy.

Bi 36
CHN ON VI NM CRYPTOCOCCUS

1. MM BNH: Cryptococcus neoformans
2. BNH PHM
Thng l dch no ty, ngoi ra c th l m, cht ht t ph qun, sinh thit c quan hoc bu.
3. QUAN ST TRC TIP
Quan st vi mc tu: ha tan mt git dch no ty hoc mt git bnh phm khc vi mt git mc
Tu ri em kho st di knh hin vi, ta nhn thy nhng t bo men d = 5 20m, chung quanh c mt
bao dy sng ng knh bng 2040m.

Cc mu sinh thit c x l theo phng php m hc; nhum HE hoc PAS di knh hin vi thy
t bo men v nang.
4. CY NM
Bnh phm cy ln mi trng Sabouraud + Chloramphenicol, 37
O
C v 25
O
C. Sau hai n ba ngy vi
nm mc to thnh khun lc nho, mu vng nu nht, kho st la cy di knh hin vi thy nhiu t bo
men c nang.

5. GY NHIM CHO TH NUI PHNG XT NGHIM
Chch bnh phm hoc la cy vo no hay mng bng ca chut bch. Khi chut cht, ly ni tng ca
chut kho st trc tip v cy ln mi trng Sabouraud thy li t bo men c nang.

CU HI LNG GI
1. M t k thut soi dch no ty vi mc tu.
2. Trnh by phng php nh danh Cryptococcus neoformans phng xt nghim.


Bi 37
VI NM HOI SINH

1. I CNG
Vi nm hoi sinh l nhm nm si hoc men c nhiu trong thin nhin, khng kh, cy c, ni m
thp,
a s nm hoi sinh khng gy bnh nhng mt s c th gy bnh c hi, gy nhim mi trng,
nhim c thc phm,
phng xt nghim c th gp cc loi bnh phm: gic mc, da, m ca bnh vim ng tai ngoi,
m.
2. HNH TH
Hnh th ca vi nm hoi sinh c xp loi da trn nhng c tnh hnh th cu trc nm:
Bo t (Conidium conidia).
Bo i (Conidiophore).
Phong bo i (Sporangiophore).
Thn si nm (Thallus).
2.1. Nhm vi nm hoi sinh da trn c tnh hnh th bo t
Alternaria sp.
Cephalosporium sp (Acremonium sp).
Curvularia sp.
Fusarium sp.
Helminthosphorium sp.
Nigrospora sp.
Scopulariopsis sp.
Monospora sp.
2.2. Nhm vi nm hoi sinh da trn c tnh hnh th bo i
Aspergillus sp.
Cladosporium sp (Hormodendrum).
Paecilomyces sp.
Penicillium sp.
Trichoderma.
2.3. Nhm vi nm hoi sinh da trn c tnh hnh th phong bo i
Absidia sp.
Mucor sp.
Rhizopus sp.
Syncephalastrum sp.
2.4. Nhm vi nm hoi sinh da trn c tnh hnh th thn si nm
Aureobasidium pullulans.
Geotrichum sp.
Rhodotorula sp.
Streptomyces sp.
3. C IM CC LOI VI NM HOI SINH
3.1. Alternaria sp
a) Khun lc nm
Mc nhanh.
Mt khm phng nh nhung.
Mu xanh l cy n nu en.
Sc t pha sau en.
b) Hnh th quan st di knh hin vi
Si t nm phn vch, mu nu en.
Bo i khng phn nhnh.
Bo t mu en, c nhiu vch ngn.

3.2. Cephalosporium sp
a) Khun lc nm
Mc nhanh.
Mt khm hi nhn, c lng t.
Mu trng xm hoc hng.

Khng sc t.
Si t nm phn vch.
Bo i phn nhnh hoc khng phn nhnh.
Bo t hnh bu dc, di, i khi phn vch
23 t bo.
Bo t thng ng thnh chm u bo i.

3.3. Curvularia sp
a) Khun lc nm
Mc nhanh.
Mt khm nh bng.
Mu nu sm n en.
Si t nm phn vch, mu nu en.
Bo i khng phn nhnh.

Bo t to, vch dy mu en, c nhiu vch ngn, hi cong.
3.4. Fusarium sp
a) Khun lc nm
Mc nhanh.
Mt khm nh bng.
Mu trng, hng hay mu xanh nht.

Si t nm phn vch.
Bo i ngn, di, phn nhnh hoc khng phn nhnh.
Bo t c nhiu vch ngn, hnh tri chui hay hnh li lim.
Cc bo t i khi ng thnh chm u bo i.
i khi bo t trn hay hnh bu dc u bo i.
3.5. Helminthosporium sp
a) Khun lc nm
Mc nhanh.
Mt khm nh bng.
Mu xm.
Si t nm phn vch, mu en.
Bo i phn nhnh hoc khng phn nhnh, i khi
c nt.
Bo t phn vch, bu dc, vch dy.

3.6. Nigrospora sp
a) Khun lc nm
Mc nhanh.
Mt khm nh len hoc bng.
Mu trng n xm.
Si t nm phn vch.
Bo i khng phn nhnh, hi phnh ra.
Bo t hnh cu en, sinh ra t u phnh
ca bo i.

3.7. Scopulariopsis sp
a) Khun lc nm
Mc nhanh.
Mt khm nh bt.
Mu trng hoc hng.
Si t nm phn vch.
Bo i phn nhnh hoc khng phn nhnh.
Bo t hnh tri chanh 2 u hi nhn.
Bo t mc t bo i, vch x x.

3.8. Monospora sp
a) Khun lc nm
Mc nhanh.
Mt khm nh bng.
Mu trng.
Si t nm phn vch.
Bo i di, ngn, khng phn nhnh.
Bo t hnh cu, vch dy, mc ra t bo i ngn st vi si t nm.
Hnh 37.6. Hnh th cu to
nm Nigrospora sp
3.9. Aspergillus fumigatus
a) Khun lc nm
Mc nhanh.
Mt khm phng nh nhung, bt.
Mu xm xanh n xanh l cy sm.
Sc t pha sau en.
Si t nm phn vch, phn nhnh.
Bo i di, u bo i phnh to thnh bu.
Ph y 1 hay 2 hng tiu bo i.
T tiu bo i sinh ra cc bo t
xp thnh chui di.

3.10. Aspergillus niger
a) Khun lc nm
Mc nhanh.
Mt khm phng nh len.
Mu trng n vng, khi gi c mu
nu sm hoc en.
T tiu bo i sinh ra cc bo t xp thnh
chui di.
3.11. Aspergillus flavus
a) Khun lc nm
Mc nhanh.
Mt khm lc u c mu vng sau i sang
mu xanh l cy.
T tiu bo i sinh ra cc bo t xp thnh chui.

3.12. Cladosporium sp (Hormodendrum)
a) Khun lc nm
Mc nhanh.
Mt khm phng hoc hi nh ln.
Mu xanh l cy n en.
Mt tri xanh l cy n en.
Bo i phn nhnh.
Bo t hnh bu dc to thnh chui
(si t nm, bo i, bo t: mu nu).

3.13. Paecilomyces sp
a) Khun lc nm
Mc nhanh.
Mt khm phng nh bt n nhung.
Mu vng nu, xm, xanh tm hay trng.
Bo i phn nhnh.
Tiu bo i c hnh thon di sinh ra
bo t bu dc, vch nhn, xp thnh chui.

3.14. Penicillium sp
a) Khun lc nm
Mc nhanh.
Mt khm phng nh bt.
Mu trng xanh n xanh da tri hoc c cc mu
khc.
Bo i phn nhnh, tiu bo i xp nh hnh bn tay
hay chi.
T tiu bo i sinh ra bo t trn xp thnh chui.

3.15. Trichoderma sp
a) Khun lc nm
Mc nhanh.
Mt khm nh len.
Mu trng n xanh l cy.
Sc t mu m n nu sm.

Bo i khng phn nhnh, tiu bo i mc ra t bo
i hay t nhnh nm.
T tiu bo i mc ra cc bo t ng thnh chm.

3.16. Mucor sp
a) Khun lc nm
Mc nhanh.
Mt khm nh len.
Mu trng n xm nu hay vng.
Si t nm ln, khng phn vch, phn nhnh.
Bo i phnh to thnh ti, bn trong c 1 li.
Cha nhiu bo t ln vch dy, nu en.

3.17. Rhizopus sp
a) Khun lc nm
Mc nhanh.
Mt khm nh len.
Mu trng, xm hay nu.
Ging Mucor nhng di chn phong bo i
c cu trc ging nh chm r.
3.18. Absidia sp
a) Khun lc nm

Mc nhanh.
Mt khm nh len.
Mu trng n xm, nu hay vng.

CU HI LNG GI
1. Phn bit hnh th hin vi 3 loi vi nm: Penicillium, Paecilomyces, Trichoderma.
2. Phn bit hnh th vi nm: Rhizopus, Mucor.

PHN BA
Ging Mucor v Rhizopus nhng chm r gi nm gia
2 chn phong bo i.
3.19. Syncephalastrum sp
a) Khun lc nm

Mc nhanh.
Mt khm nh len.
Mu trng n xm m.
Si t nm ln, phn nhnh, khng phn vch.
Phong bo i phnh to thnh ti, bn trong c 1 li.
Bo t phong l ti nh cha bo t xp nh t ngn
tay.


1. DUNG DCH BO QUN (C NH PHN)
1.1. Formol 10%
+ Formaldehyd 40% 100ml
+ Nc ct hoc NaCl 0,85% 1000ml
Formol 5%
+ Nc ct hoc NaCl 0,85% 1000ml
1.2. F
2
AM (Phenol Formol Alcool Methylene blue)

Dung dch m (dng trong 5 nm):
+ Xanh Methylene 2g
+ Cn ethylic 95
O
40ml

+ Phenol m c 8ml
+ Formol 10ml
+ Nc ct 40ml
Dung dch s dng:
+ Dung dch m 1ml
+ Formol 10ml
+ Phenol 0,1ml
+ Nc ct 89ml
1.3. Dung dch MIF (Merthiolate Iodin Formol)
MIF c nh tt nht cho cc loi n bo, gm 2 dung dch:
Dung dch m MIF:
+ Nc ct 250ml
+ Dung dch Merthiolate 1/1000 200ml
+ Glycerin 5ml
Dung dch Lugol:
+ Iod 5ml
+ Kali Iodua 10ml
+ Nc ct 100ml
Trc khi dng, ly 2,35ml dung dch MIF vo 0,15ml dung dch Lugol (khng c lm ngc li).
Lng phn dng l 0,25g.
1.4. Dung dch PVA (Polyvinyl alcohol)
Cn Polyvinyl dng lm cht nh hnh, bn di dng bt.
t t 5g PVA vo dung dch sau y nhit 75
O
C.

+ Glycerol 1,5ml
+ Dung dch Schaudinn (*) 93,5ml
(*) Dung dch Schaudinn gm:
+ Dung dch bo ha HgCl
2
2 phn

+ Cn 1 phn
1.5. Dung dch SAF (Sodium acetat acetic acid formol)
Bo qun c bo nang v th hot ng n bo, trng nang ca trng bo t v bo t ca
Microsporidium.
C th dng cho c tiu bn nhum vnh vin v tp trung.
+ Sodium acetate 1,5g
+ Acid acetic lnh 2ml
+ Nc ct 92ml
2. DUNG DCH XT NGHIM PHN TRC TIP
2.1. NaCl 0,85% 0,9%
+ NaCl 8,5g
+ Nc ct cho 1000ml
2.2. Lugol 5%
Dng nhum trng, u trng, c bit l cc bo nang ca n bo.
Dung dch d tr:
+ Iod 5g
+ Kali iodua 10g
+ Nc ct cho 100ml
Pha Iod v Kali iodua cho tan, sau cho nc ct vo. Hoc cho mt t Kali iodua vo mt t nc ct,
sau cho Iod vo, ha tan ri cho ht Kali iodua v nc ct vo. Lc vo chai nu, y kn trnh nh sng.
Dung dch s dng 1% 2%:
+ Iod 1g 2g
+ Kali iodua 2g 4g
+ Nc ct 100ml
Hoc pha 1/4 dung dch d tr vi nc ct em s dng.
3. HA CHT DNG TRONG PHNG PHP TP TRUNG PHN
3.1. Nc mui bo ha d = 1,15 1,20
+ NaCl 25 30g
+ Nc ct cho 100ml
3.2. ZnSO
4
33,1%

+ ZnSO
4
bt 33,1g

+ Nc ct 100ml
3.3. Formol 10%
3.4. Ether nguyn cht
3.5. Glycerin nguyn cht
4. DUNG DCH ST TRNG
4.1. Phenol 5%
4.2. Formol 4% 5%
4.3. Nc Javel
PH LC 2. HA CHT THUC NHUM MI TRNG
TRONG XT NGHIM NM
1. HA CHT
1.1. Potassium hydroxide (KOH): 10%; 20%
+ KOH 10g hoc 20g
+ Glycerin 10ml
+ Nc ct cho 100ml
1.2. Dung dch phm xanh Lactophenol Coton Blue (LPCB)
+ Lactic acid 20g
+ Phenol 20g
+ Xanh Cotton 50mg
+ Glycerin 40g
+ Nc ct 20ml
1.3. Nc mui sinh l 8,5% hoc 0,9%
+ NaCl 8,5 hoc 9g
+ Nc ct 1000ml
1.4. Mc tu hoc Parker en N
0
51

+ Mc tu (Nigrosin bt) 10g
+ Formol 10 % 100ml
2. MI TRNG
2.1. Thch Sabouraud (Sabouraud dextrose agar)
+ Dextrose 40g
+ Peptone 10g
+ Thch 20g
+ Nc ct 1000ml
Ha tan tt c trong nc un si, chnh pH sau cng ca mi trng n 5,6. Phn phi mi trng vo
cc ng nghim c np vn, hp kh trng 118
O
C trong 10 pht. Lu gi mi trng 4
O
C 6
O
C.

2.2. Sabouraud Cycloheximide Chloramphenicol agar (Mycobiotic agar)
+ Glucose 20g
+ Peptone 10g
+ Thch 20g
+ Chloramphenicol 50mg
+ Cycloheximide (Actidione) 500mg
+ Nc ct 1000ml
un nng ha tan mi trng trong nc, cho thm Chloramphenicol, Cycloheximide, hp kh trng
118
O
C trong 10 pht.

Chloramphenicol
+ Cn ethylic 95
o
10ml

S lng ny dng cho 1 lt mi trng (0,05mg/ml) c th thay cho Chloramphenicol bng:
* Streptomycine 40 n v/ml mi trng
* Penicilline 20 n v/ml mi trng
Cycloheximide
+ Cycloheximide 500mg
+ Acetone 10ml
S lng trn dng cho 1000ml mi trng.
2.3. Thch khoai ng (Potato Dextrose Agar)
+ Khoai ty ti hoc bt 200g
+ Dextrose 10 20g
+ Nc ct 1000ml
Hm khoai ty khong 1 gi, nghin nt lc ly nc, thm nc ct cho 1000ml, thm ng v 20g
thch, un si tr li cho tan thch, phn phi ra ng nghim c np vn, hp kh trng 120
O
C trong 10
pht, chnh pH sau cng bng 5,6.
2.4. Thch bt bp (Corn Meal Agar)
+ Bt bp 50g
+ Thch 15g
+ Nc ct 1000ml
Ha tan bt bp, thch trong nc ct un si, thm 1% Tween 80 ri phn phi ra ng nghim, em
hp kh trng 120
O
C trong 10 pht.

Ghi ch: phn bit Trichophyton rubrum v Trichophyton mentagrophytes da vo s thnh lp sc
t (Trichophyton rubrum sinh sc t ), ngi ta dng mi trng ny nhng thay th 1% Tween 80
bng 1% Glucose.
2.5. Thch tim c hm (Brain Heart Infusion = BHI)
+ BHI 40g
+ Thch 15g
+ Nc ct 1000ml
Ha tan tt c trong nc, un si, hp kh trng 120
O
C trong 15 pht, ngui 40 50
O
C, cho thm
50ml mu "O", sau phn phi ra ng nghim hoc a.
2.6. BHI c Chloramphenicol v Cycloheximide
+ BHI 37g
+ Cycloheximide 500mg
+ Thch 20g
+ Nc ct 1000ml
un nng ha tan BHI, thch, thm Chloramphenicol v Cycloheximide, hp kh trng 118
o
C trong
10 pht.
2.7. Mi trng cm (mi trng cho hoa)
+ Go trng 8g
+ Nc ct 25ml
Cho go trng v nc ct vo trong ng nghim ln hoc trong bnh Erlenmeyer y np. Nu chn.
Hp kh trng 15 18 pht. ngui. Lu gi t lnh 4 6
O
C. Khi cy, cho mng nm cn nh danh
vo mi trng nhit phng.
3. THUC NHUM
3.1. Nhum PAS
+ Acid Periodic 5g
+ Nc ct 100ml
Pha xong trong chai c nt vn.
+ Basic fuchsin 0,1g
+ Cn ethylic 95
O
5ml

+ Nc ct 95ml
Ch nc ct vo ru cho Basic fuchsin vo n khi tan ht.
+ Hydrosulfite Zn hoc Na 1g
+ Acid tartarique 5g
+ Nc ct 100ml
+ Light green 1g
+ Cn ethylic 80
O
100ml

3.2. Nhum Gram
3.3. Nhum khng acid

TI LIU THAM KHO

1. Thc tp K sinh trng Y hc. 2004. B mn K sinh trng, Trng i hc Y H Ni.
2. Thc tp K sinh trng Y hc. 2004. B mn K sinh trng, Khoa Y, i hc Y Dc TP H Ch
Minh.
3. Th Nhun v Glenns Bulmer. 1973. Vi nm hc y khoa thc dng, Y khoa i hc ng Si
Gn.
4. Boure P. 1989. Ai demmoire de Parasitologie et de Pathologie tropicale. Mdcine Sciences
Flammarion.
5. Koneman EW, Allen S D, Janda W M, Schreckenbergerand PC and Winn WC., Jr. 1997. Color Atlas
and Texbook of Diagnostic Microbiology. 5th Edition.
6. Lynne S. Garcia. 1999. Practical guide to diagnostic parasitology. American Society for
Microbiology, Washington D.C.
7. Murray PR, Baron EJ, Jorgensen JH, Pfaller MA and Yolken RH. 2003. Manual of Clinical
Microbiology. 8th Edition. American Society for Microbiology, Washington D.C.
8. Mougeot G. 1995. Conduite des examens en parasitologie. Masson, Paris Milan Barcelone.
9. Rose NR, Hamilton RG and Detrick B. 2002. Manual of Clinical Laboratory Immunology. 6th
Edition. American Society for Microbiology, Washington D.C.
10. Wery M. 1995. Protozoologie mdicale. De Boeck Universit.
11. WHO. 2003. Manual of Basic Techniques for a Health Laboratory. 2nd Edition.
12. Hnh nh ly t INTERNET.

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
Bin tp m thut v trnh by ba:
INH XUN DNG
Ch bn:
TRNH THC KIM DUNG

GIO TRNH K SINH TRNG THC HNH

M s: 7K789Y8 DAI

In 1.300 bn (Q:79), kh 19 x 27 cm. In ti Cng ty C phn In Phc Yn.
a ch: ng Trn Ph, th x Phc Yn, Vnh Phc.
S KKH xut bn: 865 - 2008/CXB/2 - 1917/GD
In xong v np lu chiu thng 12 nm 2008


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