TIU CHUN VIT NAM

TCVN 4830-3: 2005

ISO 6888-3 : 2003
V SNH VT TRONG THC PHM V THC N CHN NU - PHNG PHP NH LNG
STAPHYLOCOCC C PHN NG DNG TNH V COAGULASE (STAPHYLOCOCCUS
AUREUS V CC LO KHC; TRN A THACH - PHN 3: PHT HEN V DNG KY THUT
M S C XC SUT LN NHT (MPN) M S LNG NH
Microbiology of food and animal feeding stuffs - Horizontal method for the enumeration of coagulase-
positive staphylococci (Staphylococcus aureus and other species) - Part 3: etection and MP!
techni"ue for lo# numbers
Li gii thi!
Do tnh da dang ca thuc phm v thc n chn nui nn phuong php ny c th khng thch hop
dn tng chi tit cho tng sn phm cu th. Trong trung hop ny, c th s dung cc phuong php
khc dc trung cho tng sn phm, nu hon ton chi v l do k thut. Tuy nhin, cn c gng p
dung phuong php ny khi c th.
Khi tiu chun ny duoc sot xt tip th cn phi tnh dn moi thng tin lin quan dn pham vi m
phuong php dm da ny phi tun theo v cc nguyn nhn gy sai lch so vi phuong php trong
trung hop cc sn phm cu th.
Vic hi ho cc phuong php th c th khng thuc hin duoc ngay v di vi mt vi nhm sn
phm c th tn tai cc tiu chun quc t v/hoc tiu chun quc gia m khng ph hop vi tiu
chun ny. Trong trung hop c sn tiu chun quc t cho sn phm cn th nghim th phi tun
theo tiu chun d. Hy vong rng khi cc tiu chun nhu th duoc sot xt, th chng phi duoc sa
di d ph hop vi tiu chun ny, sao cho cui cng chi cn cc sai lch vi phuong php dm da
ny l cc l do k thut duoc tha nhn.
VI SINH V"T T#ON$ TH%C &HM V' TH(C )N CH)N NU*I - &H+,N$ &H-& ./NH L+0N$
STA&H1LOCOCCI C2 &H3N (N$ 4+,N$ T5NH V6I COA$ULAS7 8STA&H1LOCOCCUS
AU#7US V' C-C LO'I 9H-C: T#N .;A TH<CH - &H=N 3: &H-T HIN V' 4>N$ 9? THU"T
.@M SA C2 B-C SUCT L6N NHCT 8M&ND .E .@M SA L+0N$ NHF
Microbiology of food and animal feeding stuffs - Horizontal method for the enumeration of
coagulase-positive staphylococci (Staphylococcus aureus and other species) - Part 3:
etection and MP! techni"ue for lo# numbers
G &hHI Ji KL MNOg
Tiu chun ny qui dnh phuong php pht hin v dnh luong staphylococci c phn ng duong
tnh vi coagulase trn da thach ng k thut dm s c xc sut ln nht (MPN). Tiu chun ny
c th p dung cho:
- Cc sn phm dng cho con ngui v thc n chn nui, v
- Cc mu mi trung trong khu vuc sn xut v x l thuc phm.
Tiu chun ny duoc khuyn co p dung cho cc sn phm khi uc tnh tnh staphylococci c mt
trong sn phm vi s luong nh, v du nhu cc sn phm kh. Staphylococci c phn ng duong
tnh vi coagulase an du l cc Stapylococcus aureus nhung Stapylococcus intermedius v mt s
chng ca Stapylococcus hyicus cng sinh ra coagulase.
2 TPi Qi! JiO MRO
Cc ti liu vin dn sau l rt cn thit cho vic p dung tiu chun. i vi cc ti liu vin dn ghi
nm an hnh th p dung phin n duoc nu. i vi cc ti liu vin dn khng ghi nm an hnh
th p dung phin n mi nht, ao gm c cc sa di.
TCVN (SO ) (tt c cc phn), Vi sinh vt trong thuc phm v thc n chn nui - Chun
mu th, huyn ph an du v cc dung dch pha long thp phn d kim tra vi sinh vt.
TCVN 3 (SO ), Sa v cc sn phm sa. Chun mu th v cc dung dch pha long d
kim tra vi sinh.
TCVN 3- : (SO - : , ith amendment : 3), Vi sinh vt trong thuc phm v
thc n chn nui - Phuong php dnh luong staphylococci c phn ng duong tnh vi coagulase
(Staphylococuss aureus v loi khc) trn da thach. Phn : K thut s dung mi trung thach
aird-Parker.
TCVN 3- : (SO - : , ith amendment : 3), Vi sinh vt trong thuc phm v
thc n chn nui - Phuong php dnh luong staphylococci c phn ng duong tnh vi coagulase
(Staphylococuss aureus v loi khc) trn da thach. Phn : K thut s dung mi trung thach
irinogen huyt tuong th.
TCVN (SO ), Vi sinh vt trong thuc phm v trong thc n chn nui - Nguyn tc chung
d kim tra vi sinh vt.
SO/TS 33-, Microiology o ood and animal eeding stus - Guidelines on preparation and
production o culture media - Part : General guidelines on quality assurance or the preparation o
culture media in the laoratory (Vi sinh vt trong thuc phm v thc n chn nui - Hung dn chun
v tao mi trung cy - Phn . Cc hung dn chung v dm o cht luong cho vic chun
mi trung cy trong phng th nghim).
SO/TS 33- ; 3, Microiology o ood and animal eeding stus - Guidelines on preparation
and production o culture media - Part : Practical guidelines on perormance testing o culture media
(Vi sinh vt trong thuc phm v thc n chn nui - Hung dn chun v tao mi trung cy -
Phn : Cc hung dn thuc hnh v kim tra tnh nng ca mi trung cy).
3 Th!St OgT JP UVOh OghWX
Trong tiu chun ny s dung cc thut ng v dnh ngha sau dy:
3YG
staphylococci c phn ng duong tnh vi coagulase (coagulase -positive staphylococci) vi khun
hnh thnh cc khun lac din hnh v/hoc khng din hnh trn mt mi trung cy chon loc v
cho cc phn ng duong tnh vi coagulase hoc phn ng huyt tuong th dc trung trn thach
irinogen huyt tuong th.
CH THCH: Tiu chun ny khng dnh staphylococci c phn ng duong tnh vi coagulase duoc
dua trn phn ng duong tnh vi coagulase manh, nhung phi cng nhn rng mt s chng
staphylococci c phn ng duong tnh vi coagulase nhung cho cc phn ng duong tnh vi
coagulase yu. Cc chng ny c th nhm vi cc vi khun khc nhung chng c th duoc phn
it ng cc th nghim sung nhu vic tao ra thermonucleasa (v chi tit, xem D 3).
3Y2
.VOh QZ[Og \tXLh]Q^_^__i _` LhaO bOg MZcOg tdOh Ji _^Xg!QX\e (enumeration o coagulase-
positive staphylococci)
Vic xc dnh s luong staphylococci c phn ng duong tnh vi coagulase tm thy trong mt gam
hoc mt mililit mu khi tin hnh th nghim theo phuong php qui dnh trong tiu chun ny.
4 Ng!]fO tg_
4YG &hZcOg LhKL LhKt hiO
4YGYG Cy ln mi trung chon loc mt luong mu th qui dnh, nu sn phm an du dang lng,
hoc mt luong huyn ph qui dnh an du nu cc sn phm dang khc .
4YGY2 cc ng trong mi trung k kh 3
o
C t h dn h. Su c mt ca staphylococci gi
dnh c phn ng duong tnh vi coagulase duoc chi th i su kh kali telurit.
CH THCH: Trong tiu chun ny, mi trung k kh thu duoc ng cch rt thach hoc parain
dy ln pha trn mi ng, nhung cng c mt cch khc l cc ng ny trong nh hoc trong t
m trong cc diu kin k kh.
4YGY3 Cy ln mt mi trung aird-Parker dc chon loc cc khun lac ly t cc ng duong tnh
gi dnh (..) sau h v tt c cc ng cn lai sau h.
4YGY4Y Tt c cc ng duoc cy 3
o
C t h dn h. Su c mt ca staphylococci gi dnh c
phn ng duong tnh vi coagulase duoc chi th i su kh kali telurit v phn ng vi lng d
trng.
4YGY5 Cc khun lac din hnh v/hoc khng din hnh duoc khng dnh ng phn ng vi
coagulase.
4YGY6 Cch khc, c th cy ln mt thach irinogen huyt tuong th, sau khi m thch hop, su
c mt ca staphylococci c phn ng duong tnh vi coagulase duoc chi th i cc khun lac cho
thy c phn ng vi irinogen huyt tuong th dc trung.
4YGYh Kt qu l c mt hay khng c mt staphylococci c phn ng duong tnh vi coagulase
trong x g hay x ml sn phm.
4Y2 &hZcOg LhKL UVOh QZ[Og
4Y2YG Cc dy dung dch pha long sn phm duoc cy vo mi trung cy lng chon loc.
4Y2Y2 Cc ng duoc trong diu kin k kh 3
o
C t h dn h. Su c mt ca staphylococci
c phn ng duong tnh vi coagulase gi dnh duoc chi th i su kh kali telurit.
CH THCH: Trong tiu chun ny, mi trung k kh thu duoc ng cch rt thach hoc parain
dy ln pha trn mi ng, nhung cng c mt cch khc l cc ng ny trong nh hoc trong t
m trong cc diu kin k kh.
4Y2Y3 Cy ln mt mi trung aird-Parker dc chon loc cc khun lac ly t cc ng duong tnh
gi dnh (..) sau h v tt c cc ng cn lai sau h.
4Y2Y4 cc ng ny 3
o
C t h dn h. Su c mt ca staphylococci gi dnh c phn ng
duong tnh vi coagulase duoc chi th i su kh kali telurit v phn ng vi lng d trng.
4Y2Y5 Cc khun lac din hnh v/hoc khng din hnh duoc khng dnh ng phn ng vi
coagulase.
4Y2Y6 Cch khc, c th cy ln mt thach irinogen huyt tuong th, sau khi m thch hop, su
c mt ca staphylococci c phn ng duong tnh vi coagulase duoc chi th i cc khun lac cho
thy c phn ng vi irinogen huyt tuong th dc trung.
4Y2Yh S c xc sut ln nht ca staphylococci c phn ng duong tnh vi coagulase trong mt
gam hay trong mt mililit sn phm tnh duoc ng cch di chiu vi cc ng s c xc sut ln
nht cho cc d pha long khng dnh ( . hoc ..)
5 4V_h LhX Q^iOg JP Iji tkZOg _l]
i vi phng th nghim, xem TCVN (SO ).
Cc ha cht duoc s dung d chun mi trung cy v thuc th phi c cht luong tinh khit
phn tch.
5YG 4V_h LhX Q^iOg
Xem phn tuong ng ca TCVN (SO ), hoc TCVN 3 (SO ), hoc tiu chun
ring lin quan dn sn phm cn kim tra.
5Y2 Mji tkZOg $i^Qitti JP CXOt^Oi _ai minO
5Y2YG Mji tkZOg _c maO
5Y2YGYG ThPOh LhoO
Mi trung nng d
kp
Mi trung nng d
don
Pepton t casein , g , g
Cao tht , g , g
Cao men , g , g
Liti clorua , g , g
Mannitol , g , g
Natri clorua , g , g
Glyxin , g , g
Natri pyruvat , g 3, g
Polyoxyetylen soritan mono-oleat (Teen
)
, g , g
Nuc ml ml
5Y2YGY2 Ch!pO mV
Ho tan cc thnh phn trong nuc, ng cch dun nng v khuy, nu cn, d thu duoc dung dch
ho tan hon ton. Lm ngui dn nhit d phng v chinh pH sao cho sau khi kh trng pH cui
cng l , ,, nu cn.
Phn phi mi trung vi cc luong thch hop vo cc ng nghim c kch thuc ph hop (v du: di
vi mi trung nng d don, dng ng mm x mm, di vi mi trung nng d kp th dng
ng mm x mm).
Kh trng pht trong ni hp p luc
o
C.
5Y2Y2 4!Og MV_h qXQi teQ!kit
5Y2Y2YG ThPOh LhoO
Kali telurit
)
(KTeO3) , g
Nuc ml
5Y2Y2Y2 Ch!pO mV
Ho tan kali telurit trong nuc ng cch dun nng rt nh.
t phi d tan. Nu c mt cht khng tan mu trng trong nuc; th loai kali telurit.
Loc qua mng loc c l , m d kh trng.
Dung dch c th o qun duoc ti da mt thng nhit d 3
o
C
o
C.
Loai dung dch nu c kt ta mu trng.
5Y2Y3 Mji tkZOg h^PO _hrOh
Ngay truc khi s dung, dun nng mi trung co n (..) pht
o
C d dui ht khng kh.
Lm ngui dn nhit d t
o
C dn
o
C v ng k thut v trng cho thm vo mi ng dung
dch kali telurit (..): , ml di vi mi trung nng d don v , ml di vi mi trung nng d
kp.
5Y2Y4 Ths tdOh OtOg Uu UaI ma^ _hlt QZ[Og Iji tkZOg _l]
xc dnh tnh chon loc v nng sut, xem SO/TS 33-. ng cho thy chun c th nghim
tnh nng ca mi trung Giolitti v Cantoni ci in.
vaOg G - Ch!pO _b ths OghiI tdOh OtOg _wX Iji tkZOg $i^Qitti JP CXOt^Oi _ai minO
Chb_
OtOg
Chn Ux w CK_ _hwOg qiuI tkX
Mji
tkZOg
_h!pO
&hZcOg
LhKL qiuI
tkX
Ch!pO _b
Nng
sut
3
o
C
trong
h
Staphylococcus aureus ATCC
3 P hoc Staphylococcus
aureus ATCC 3 cng vi
chng canh tranh ($%coli ATCC
3 hoc ), hoc chng
tuong tu duoc dng k trong cc
khc
n dnh
luong
khun
lac, trn mi
trung chon
loc
2)
Chi s dung kali telurit ph hop di vi php th ny (.3..)
Tnh
chon loc
3
o
C
trong h
$scherichia coli ATCC
hoc 3 hoc chng tuong tu
duoc dng k trong cc khc
TSA
n dnh
luong
Khng moc
trn mi
trung khng
chon loc
5Y3 Mji tkZOg thH_h ( g/l)
5Y3YG ThPOh LhoO
Thach g dn g
3)
Nuc ml
5Y3Y2 Ch!pO mV
Ho tan thach trong nuc ng cch dun si v kh trng pht trong ni hp p luc
o
C.
Lm ngui dn nhit d t
o
C dn
o
C truc khi s dung. Rt vo cc ng nghim c dung tch
thch hop. o qun theo TCVN (SO ).
5Y4 Mji tkZOg thH_h vXikM-&Xkqek
5Y4YG ThPOh LhoO JP _h!pO mV
Xem .3 ca TCVN 3- : (SO - : , ith amendment : 3).
5Y4Y2 Ths tdOh OtOg Uu UaI ma^ _hlt QZ[Og Iji tkZOg _l]
xc dnh tnh chon loc v nng sut, xem SO/TS 33-. i vi cc chun c, xem ng .
ca SO/TS 33-.
5Y5 Mji tkZOg thH_h yimkiO^geO h!]nt tZcOg thz 8{eI |6} JP |h}D
5Y5YG ThPOh LhoO JP _h!pO mV
Xem .3 ca TCVN 3- : (SO -: , ith amendment : 3).
5Y5Y2 Ths tdOh OtOg Uu UaI ma^ _hlt QZ[Og Iji tkZOg _l]
xc dnh tnh chon loc v nng sut, xem SO/TS 33-. i vi cc chun c, xem ng .
ca SO/TS 33-.
5Y6 Mji tkZOg _XOh thXOg Oi^ - tiI 8vkXiO-heXkt iOy!\i^O mk^thD
5Y6YG ThPOh LhoO JP _h!pO mV
Xem . ca TCVN 3- : (SO -: , ith amendment : 3).
5Y6Y2 Ths tdOh OtOg Uu UaI ma^ _hlt QZ[Og Iji tkZOg _l]
xc dnh tnh chon loc v nng sut, xem SO/TS 33-. i vi cc chun c, xem ng .
ca SO/TS 33-.
5Yh H!]nt tZcOg thz
Xem . ca TCVN 3- : (SO -: , ith amendment : 3).
6 Thint mV JP MNOg _N
CH THCH: C th dng dung cu thu tinh s dung mt ln thay th cho cc dung cu thy tinh s
dung nhiu ln nu chng c cc dc tnh thch hop.
S dung cc thit , dung cu ca phng th nghim vi sinh thng thung [xem TCVN (SO
)] v cu th l:
6YG Tw lI, c th hoat dng 3
o
C
o
C.
6Y2 Tw \l] hoc Q~ \l] UZ[_ thng gi ng di luu, c th duy tr nhit d t 3
o
C dn
o
C. C
th s dung t quat gi.
6Y3 .WX &etki, v trng.
3)
Ty thuc vo sc dng ca thach
6Y4 !e _l] J~Og, ng platin-iridi, niken-crm hoc cht do, dung knh khong 3 mm, hoc cc
que cy vng l v trng s dung mt ln v _K_ qiI _l] \! ng cht liu tuong tu.
6Y5 AOg OghiI, c dung tch thch hop (v du: mm x mm, mm x mm v mm x
mm).
6Y6 &iLet _hiX Ux, c dung tch danh dnh ml, duoc chia vach , ml v c l x vi dung knh
hop l.
6Yh Ni _K_h thw], c th hoat dng nhit d t
o
C dn
o
C v 3
o
C.
6Y8 4X^, d ct thach.
6Y vOh q qhdY
h Ll] IR!
iu quan trong l phng th nghim nhn duoc dng mu dai din v khng hu hng hoc in
di trong sut qu trnh vn chuyn v o qun.
Vic ly mu khng qui dnh trong tiu chun ny. Nu chua c tiu chun ring v ly mu cho sn
phm tuong ng, th cc n c lin quan tu tho thun v vn d ny.
8 Ch!pO mV IR! ths
Vic chun mu th theo cc phn thch hop ca TCVN (SO ), hoc TCVN 3 (SO
) hoc tiu chun ring cho sn phm tuong ng. Nu chua c tiu chun ring th cc n lin
quan tu tho thun vi nhau v vn d ny.
CK_h tinO hPOh
YG &hZcOg LhKL LhKt hiO
YGYG &hoO IR! ths JP h!]O Lh mXO Uo!
Xem cc phn tuong ng ca TCVN (SO ) tuy thuc vo sn phm c lin quan hoc
TCVN 3 (SO ).
chun huyn ph an du, cho ml hoc g sn phm vo ml hoc g mi trung Giolitti v
Cantoni ci in nng d don (.) hoc ml hoc g vo ml hoc g mi trung Giolitti v
Cantoni ci in nng d kp. i vi cc th tch ln hon ca cc phn mu th, thm x ml hoc x g
vo x ml hoc x g mi trung Giolitti v Cantoni ci in nng d don d duoc dui kh v d
duoc sung kali telurit, v c th tch khng kh nh nht trong nh hoc trong vt cha. Rt cn
thn thach (.3) hoc parain ph ln trn mi trung, duoc lm ngui dn nhit d t
o
C dn
o
C v cho dng dc lai d tao thnh lp hn kn.
YGY2 TtOg \iOh
(xem .) dung dch huyn ph an du (..) trong h h 3
o
C. Nu den hoc c kt
ta den th cy truyn nhu trong ..3. Nu khng hin mu den th tip h h v cy truyn
nhu trong ..3 (k c xut hin hoc khng xut hin mu den hoc kt ta den).
YGY3 Cl] tk!]O t _K_ Og ttOg \iOh
ng cch v trng, tho lp thach hoc parain pha trn, s dung dao v trng (.) d ct (xem
ch thch trong ..) thnh n phn theo chiu doc. Nu cn, chn dao vo st thnh theo dung
cong ca ng thu tinh d tch lp ph ny. Lc manh ng d lp ph v thnh mnh nh roi xung
dy ng v dm o cn nguyn huyn ph dch cy.
Dng que cy vng v trng (.), ly mt vng dy tng mi trung chon loc dn ln mt cc da
thach aird Parker (.) ring it hoc cc da thach irinogen huyt tuong th (.) d thu duoc
cc khun lac phn lp.
Lt nguoc cc da d chun v dt chng vo t m (.) d 3
o
C trong h h v h
h.
Y2 &hZcOg LhKL UVOh QZ[Og
Y2YG &hoO IR! ths JP h!]O Lh mXO Uo!
Xem cc phn thch hop ca TCVN (SO ) tuy thuc vo sn phm c lin quan hoc
TCVN 3 (SO ).
Y2Y2 Cl]
Ly a ng mi trung nng d kp, d dui kh v d sung kali telurit (..3). Chuyn vo mi
ng ml mu th dang lng hoc ml dung dch pha long an du (tc l g mu) ca cc sn
phm dang khc.
Ly a ng mi trung nng d don, d dui kh v sung kali telurit (..3). Chuyn vo mi ng
ml mu th dang lng hoc ml dung dch pha long an du (tc l , g mu) ca cc sn phm
dang khc.
i vi mi d pha long tip theo (tc l
-
,
-
v
-3
di vi sn phm dang lng hoc
-
,
-3

hoc
-
di vi cc sn phm dang khc), tin hnh nhu m t trn, s dung pipet v trng mi
cho mi d pha long.
Chun d mt s luong cc dung dch pha long d dm o rng d pha long cui cng d d
cho a kt qu m tnh.
Trn cn thn cht cy v mi trung, trong mi trung hop trnh d ln khng kh vo.
Cn thn rt mt lp thach (.3), d duoc lm ngui dn nhit d t
o
C dn
o
C ln trn mi
trung trong mi ng d cy v d cho dc lai tao thnh lp hn kn.
Y2Y3Y
(xem .) cc ng nghim dung mi trung nng d kp v mi trung nng d don (..) d
cy, 3
o
C trong h h. Cy truyn cc ng nghim cho thy den hoc c kt ta den nhu d
chi ra trong ..
tip cc ng cn lai trong h h v cy truyn tt c cc ng nhu trong ..3 (k c cc ng c
kt ta den hoc khng c kt ta den sau h h).
Y2Y4Y Cl] tk!]O
Y3 ChO UWX Ja giai thd_h qnt !a
Y3YG Mji tkZOg thH_h vXikM-&Xkqek
Y3YGYG ChO _K_ qh!pO QH_
Sau khi cc da (..3 hoc ..) h, dnh du vo dy cc da cc v tr ca t ky khun lac
din hnh no c mt.
CH THCH : Cc khun lac din hnh c mu den hoc mu xm, ng v li (dung knh t mm
dn , mm sau khi trong h, v c dung knh t , mm dn , mm sau khi h) v duoc
ao quanh ng mt vng trong r rt, cng c th l m tng phn. Sau khi trong t nht h, c
th xut hin mt vng mu trng duc tip gip vi khun lac .
CH THCH : Cc khun lac khng din hnh cng kch c nhu khun lac din hnh v c th c
mt trong cc hnh thi sau.
a) Cc khun lac den ng c hoc khng c ra trng hp; khng c vng trong hoc hu nhu khng
nhn thy v khng c vng trng duc hoc rt kh nhn thy.
) Cc khun lac mu xm khng c vng trong.
Cc khun lac khng din hnh duoc hnh thnh ch yu do cc chng staphylococci c phn ng
duong tnh vi coagulase nhim vo sn phm, v du: cc sn phm sa, tm v cc phn ca
vt nui. Chng thung t khi duoc hnh thnh do cc chng staphylococci c phn ng duong tnh
vi coagulase nhim trong cc sn phm khc.
CH THCH 3: Cc khun lac khc l tt c cc khun lac cn lai c kh nng c mt trn cc da m
khng cho thy iu hin n ngoi din hnh hoc khng din hnh nhu d m t trong Ch thch
v Ch thch , v duoc coi l h vi khun nn.
lai (xem .) tt c cc da 3
o
C thm h + h v dnh du t ky khun lac din hnh no
mi. ng thi cng dnh du t ky khun lac khng din hnh no c mt.
Y3YGY2 9hOg UVOh
T mt ca mi khun lac d chon (.3..), dng kim cy v trng ly mt phn v chuyn vo
ng hoc lo dung mi trung canh thang no-tim (.).
(xem .) 3
o
C trong h h.
ng k thut v trng, ly , ml chng cy cho vo ,3 ml huyt tuong th (.) (tr khi nh sn
xut qui dnh cc luong khc) dung trong cc ng v trng c kch thuc thch hop (v du mm x
mm)
Nghing ng, kim tra su kt dnh ca huyt tuong sau khi t h dn h, v nu php th l m
tnh th kim tra lai sau khi h h, hoc kim tra theo thi gian duoc nh sn xut qui dnh.
Cc php th coagulase duoc coi l duong tnh nu cc chng cy sinh ra tai t nht cc phn ng
coagulase 3
+
theo hung dn dnh du (k vach) trong Hnh . Cc phn ng t
+
dn
+
duoc coi
l trung gian.
V kim tra m tnh, di vi mi m huyt tuong, thm , ml mi trung canh thang no-tim v
trng (.) vo mt luong huyt tuong th (.) d duoc khuyn co v nhung khng cy. Vic
kim tra huyt tuong khng c du hiu no ca su kt dnh th php th l hop l.
nh du duong tnh mi ng c t nht mt khun lac duoc khng dnh l duong tnh vi
coagulase.
- m tnh: khng c ng chng hnh thnh irrin.
+ duong tnh: kt dnh nh khng c t chc
+ duong tnh: kt dnh nh c t chc
3 + duong tnh: kt dnh nhiu c t chc
+ duong tnh: ton kt dnh v khng dch chuyn khi ng lt nguoc
HOh G - .KOh Ml! _K_ LhaO bOg ths OghiI Ji _^Xg!QX\e
Y3Y2 Mji tkZOg thH_h yimkiO^geO h!]nt tZcOg thz
Sau khi h h, v tip h nu cn, staphylococci c phn ng duong tnh vi coagulase
hnh thnh cc khun lac nh mu den hoc mu xm (hoc thm ch mu trng) duoc ao quanh
i vng sng kt ta du hiu cho hoat tnh coagulase. Khi t du , cc khun lac Proteus, c th
cho thy v n ngoi ging vi cc khun lac staphylococci c phn ng duong tnh vi coagulase.
Tuy nhin sau khi h hoc h chng c th pht trin thnh khun lac moc rng c mu nu
dm hoc nu nhat do d c th phn it duoc vi vi khun staphylococci.
Php th l duong tnh khi c mt t nht mt khun lac chi th hoat tnh coagulase duoc thy
CH THCH: Vi thach irinogen huyt tuong th duoc dua trn phn ng coagulase, nn khng cn
thit phi khng dnh hoat tnh ny.
G0 viu! thV qnt !a
G0YG &hZcOg LhKL LhKt hiO
Theo gii thch cc kt qu, ghi lai su c mt hoc khng c mt staphylococci c phn ng duong
tnh vi coagulase trong phn mu th, theo khi luong tnh ng gam hoc theo th tch tnh ng
mililit ca mu th.
G0Y2 &hZcOg LhKL UVOh QZ[Og
G0Y2YG ChO _K_ Ux LhX Q^iOg
CH THCH: Huyn ph an du v mu th, nu dang lng duoc coi l dung dch pha long.
i vi mi d pha long d duoc cy vo mi trung cy lng chon loc (.. v ..), ghi lai s
luong ng nghim cho thy c mt staphylococci c phn ng duong tnh vi coagulase d duoc
khng dnh ng php th thach aird-Parker (.3.) hoc su c mt cc khun lac vi cc phn ng
duong tnh vi coagulase trn mi trung thach irinogen huyt tuong th (.3.). Chi r d l cc
ng duong tnh.
Chon a d pha long lin tip theo . ca TCVN : (SO : ) v xc dnh chi s
c xc sut ln nht (MPN) [xem TCVN (SO )]
G0Y2Y2 TdOh t^KO
Xem . ca TCVN : (SO : ).
GGY .x _hNI
Nhu d it, cc sai lch ln trong cc kt qu c th xy ra vi k thut MPN. Cc gii han tin cy
qui dnh duoc dua hon ton trn su phn ngu nhin cc kt qu. Cc nguyn nhn gy ra sai
lch khc di khi c th rt quan trong, nhung khng d cp trong k thut MPN. tnh dn cc nh
hung ny, s dung cc pham tr trong ng . ca TCVN : (SO : ). Chng
tng kt cc t hop ng theo kh nng xut hin ca chng.
CH THCH: Mt nghin cu cng tc quc t (xem [3]) trn cc mu sa kh cho thy, s dung k
thut MPN ging nhu m t trong tiu chun ny, th trong % cc trung hop chnh lch gia cc
kt qu ca hai php th dc lp nh hon , ln so vi trung nh cng ca hai kt qu. Chnh
lch ln nht quan st duoc gia cc kt qu ca hai php th dc lp l . ln so vi trung nh
cng ca hai kt qu.
G2 vK^ _K^ ths OghiI
o co th nghim phi chi r:
a) Moi thng tin cn thit d nhn it dy d v mu th,
) Phuong php ly mu d s dung, nu it;
c) Phuong php th nghim d dng (phuong php pht hin hoc phuong php dnh luong; mi
trung d s dung), vin dn tiu chun ny;
d) Moi chi tit thao tc khng duoc quy dnh trong tiu chun ny hoc nhng diu duoc coi l tuy
cng nhu cc su c t ky m c th nh hung dn kt qu;
e) kt qu thu duoc, chi r phuong php iu th d s dung.
TH+ MC T'I LIU THAM 9H3O
[] D 3 : , Milk and milk-ased products - Detection o thermonuclease produced y
coagulase - positive staphylococci in milk and milk-ased products.
[] D C : , Milk and milk-ased products - Detection o coagulase - positive staphylococci -
Most proale numer technique.
[3] CHOPN, A. , MALCOLM, S. , JARVS, G. , ASPERGER, H. , ECKERS, H.J. , ERTONA, A.M. ,
COMNAZZN, C. , CARN, S. , LOD, R. . HAHN, G. , HEESCHEN, W. , JANS, JERVS, D.l. ,
LANER, J.M. , OCONNOR, ., REA, M. , ROSS, J. , SELGMANN, R. , TESONE, S. . WAES, G. ,
MOCQUOT, G. and PVNCK H Comparision o our media and methods or enumerating
Staphylococcus aureus in podered milk: CMS methods studies: XV, J. ood. Prot. , , , pp.
-.
[] ARD - PARKER, A.C. improvide diagnostic and selective medium or isolating coagulase
positive staphylococci, J. Applied acteriology, (), , pp. -.
[] SMTH, .A and ARD - PARKER, A.C. The use o sulphamezathine or inhiiting Proteus spp.
on aird-Parker's isolation medium or Staphylococcus aureus. J. Applied acteriology, (). ,
pp. - .
[] ECKERS, H.L et al. Evaluation o pour-plate system ith rait plasma-ovine irinogen agar or
the enumeration o Staphylococcus auraus in ood. Can. J. Microioi. , 3, , pp. -.
[] SAWHEY. D. The toxicity o potassium tellurite to Staphylococcus aureus in rait plasma
irinigen agar, J. Applied acteriology, , , pp, - .