—| iO < jz TIEU CHUAN VIET NAM TCVN 11039-6:2015 Xudt ban tn 1 PHU GIA THU’C PHAM - PHU'O'NG PHAP PHAN TICH VI SINH VAT - PHAN 6: PHAT HIEN VA DINH LUQNG STAPHYLOCOCCUS AUREUS BANG KY THUAT BEM KHUAN LAC Food aditive - Microbiological analyses - Part 6:Detection and enumeration of staphylococcus aureus by colony count technique HA NOI - 2045TCVN 11039-6:2015 Loi néi dau TCVN 11039-6:2015 duge xay dyng trén co sé tham khdo JECFA 2006, Combined compendium of food additive specifications, Volume 4: Analytical methods, test procedures and laboratory solutions used by and referencad in the food additive specifications; TCVN 11039-6:2015 do Ban kj thuat tiéu chudn TCVN/TC/F4 Gia vj va phy gia thy phdm bién soan, Téng eye Tiéu chuan Bo Ivéng Chat lveng thdm dinh, B6 Khoa hoc va Céng nghé cong bé; B6 tigu chudn TCVN 11039 Phy gia thye phém — Phuong phép phan tich vi sinh vét gdm cdc phan sau: — TCVN 11039-1:2015, Phan 1: X4c dinh téng s6 vi sinh vat hiéu khi bang ky thudt dém dia; TCVN 11039-2:2015, Phan 2: Xéc dinh téng s6 vi sinh vat hiéu khi bang ky thuét dém dia xodn; 1 TCVN 11039-3:2015, Ph&n 3: Phat hién va dinh lugng coliform va E. coli bang ky thuat dém 36 06 xéc sudt ion nhat (Phuong phép chudn); TCVN 11039-4:2015, Phan 4: Phat hién va dinh Ivong coliform va E. coli bang ky thuat dém sé 06 xe sudt ion nhét (Phuong phép théng dung); TCVN 11039-5:2015, Phan 5: Phat hién Salmonella; TCVN 11039-6:2015, Phdn 6: Phat hi6n va dinh long Staphylococcus aureus bang ky thuat dém Khuén lec; TCVN 11039-7:2015, Phdn 7: Phat hién va dinh lrgng Staphylococcus aureus bing kj thuat dim 86 6 xéc sudt lon nhét (MPN); TCVN 11039-8:2015, Phan 8: Binh iugng ném men va ném méc.TIEU CHUAN QUOC GIA TCVN 11039-6:2015 Phu gia thyc phdm - Phuong phap phan tich vi Phan 6: Phat hién va dinh lwong Staphylococcus aureus bang kj thuat dém khuan lac inh vat - Food additives - Microbiological analyses - Part 6: Detection and enumeration of Staphylococcus aureus by colony count technique 4 Pham vi 4p dung Tieu chudn nay quy dinh phuong phdp phat hign va dinh ugng Staphylococcus aureus trong phy gia thy phim bang ky thuat dém khudn lac. Phuong phap nay thich hyp 48 phan tich mau dy kién chtra trén 100 té bao S. aureus trén gam. Néu nghi ngo sé long té bao th4p hon gigi han néu trén thi nén sir dyng phuong phap dém s6 c6 xéc sudt lon nhdt theo TCVN 11039-7:2015. 2. Tai ligu vign dan Céc tai ligu vién dan sau la rt cn thiét cho viéc ap dyng tiéu chudn. 6i véi cdc tai ligu vién din ghi nm céng bé thi p dung phién ban duge néu. Bi v6i cdc tai ligu vién din khéng ghi ném cing b6 thi &p dung phién ban méi nh&t, bao gdm ca cae sia di, TCVN 11039-7:2015, Phy gia thye phém — Phuong phép phan tich vi sinh vt - Phan 7: Xéc dinh Staphylococcus aureus bang ky thugt dém sé c6 xdc suét lon nhat 3. Thuat ngi va dinh nghia ‘Trong tiéu chun nay sir dung thuat ngér va dinh nghia sau day’ 34 Staphylococcus aureus (Staphylococcus aureus) ‘Vi khudn hinh thanh khudn lac dién hinh va/hoac khéng dién hinh trén bé mat ca mdi trong cdy chon Igc va cho phan ting coagulase duong tinh khi tién hanh thir nghiém theo phuong phap quy dinh trong tigu chuan nay.TCVN 11039-6:2015 4 Nguyén tic 4.1 Céy lén bé mat cia méi truéng chon loc mét Iugng huyén phi: ban dau quy dinh. Trong cing mét diéu kién, cdy céc dung dich pha loang thap phan cla huyén phi ban dau. 4.2 U cdc dla trong didu kién hiéu khi & 35 °C va kiém tra sau thdi gian tr 18 h dén 24 h. 4.3 Tinh sé twong Staphylococcus cé phan Ung coagulase dong tinh trong mot gam mau tir sé tung khuan lac dién hinh va/ hoac khéng dién hinh trén cac dia & cac 4} pha long da chon sao cho két qua cd y nghia va due khang dinh bang két qua thy coagulase dong tinh. 5 Méi trang cay, dich pha loang va thude thir 5.1 Méi trwe@ng Baird-Parker (xem A. 1). 5.2 Canh thang trypticase (tryptic) dau twong (TSB), chiva natri clorua 10 % va natri pyruvat 1% (xem A.2). 5.3 Thach trypticase (trypton) dau twong (TSA) (xem A 3). 6.4 Thach toluidin xanh-axit deoxyribonucleic (ADN) (xem A.4). 5.5 Canh thang phenol dé cacbohydrat (xem A.5). 5.6 Coagulase plasma (tir thd) chira EDTA, v6 triing (cé ban sn) 5.7 Dung dich tysostaphin (xem A.6). 5.8 Dung dich hydro peroxit, 3 % thé tich (xem A.7). 5.9 Dung dich dgm musi phosphat, 0,02 M, chia natri clorua 1%. 5.10 Daw parafin, v6 tring. 6 Thiét bj va dyng cy thuy tinh C6 thé ding dung cy thuy tinh st dying mét lan thay thé cho c&c dyng cy thuy tinh stv dung nhidu lan néu ching ¢6 cc ac tinh thich hep. ‘Sir dung cc thiét bj ciia phdng thy nghiém vi sinh va cy thé 1a: 6.4 Budng sdy, 6 ddng khi méng ho&e phdng duge théng khi tét, khong 6 bui va dy kién mat 46. vi sinh vat trong khéng khi & khu vyc lam viéc khéng ln hon 15 khudn lac trén mdi dia sau 15 min phoi nhiém.6.2 Bia Petri, bang chat déo (15 mm x 90 mm) hoac thiy tinh (15 mm x 100 mm). 6.3 Pipet, c6 thé phan phdi 1, 5 va 10 ml, chia vach dén 0,1 ml 6.4 Ti dm, c6 thé duy tri nhiat 66 35 °C + 1 °C. 6.5 Que cdy gat, bang thiy tinh, dvdng kinh ty 3 mm dén 4 mm, dai tir 15 cm én 20 cm, 6 mot au dugc udn cong mét doan dai tty 45 mm dén 55 mm, 6.6 Thiét bj dém khudn lac, loai co hoc hodc dién tk, cé ngudn séng thich hp, dia lusi va bo dém. 6.7 Ong nghi¢m vé tring, kich thuée 13 mm x 100 mm. 6.8 Ndi céch thily, o6 thé dun dén s6i 7 Lay mau Mau giti d&n phong thi nghiém phai ld mau dai di¢n. Mau khéng bi hu héng hode bién adi trong sudt ‘qua trinh van chuyén hoae bdo quan. \Viée dy mau khong quy dinh trong tiéu chuan nay. Xem tiéu chudn cy thé o6 lién quan én san phdm. Néu chwa cé tigu chudin cy thé lién quan dén san phdm thi cdc bén cé lién quan nén thoa thugn voi nhau vé vn d& nay. 8 Chudn bj mau thir Mau dugc tron déu 48 thu duge cac phan mau thir ding nhat. 9 Cach tién hanh 9.1 Chuan bj dung dich pha loang Trong cdc diéu kién v6 tring, chun bi day dung dich pha lodng thap phan bang cach chuyén 10 ml dung dich mu thir 48 pha long true dé vao 90 mi dung dich pha loding (6.9), sir dyng cdc pipet (6.3) riéng cho ting ndng 46. Khdng lam mau nai bot. Lac t&t cd c&c dung dich pha loang 25 ln voi bien do 30 om trong vong 7 . 9.2 Phan lp D6i voi mdi dung dich pha long 48 dé dia, chuyén vé tring 1 ml vao 3 da thach Baird-Parker (5.1), phan phéi 1 ml dung dich m&u tht vao ba dia (vi dy 0,4 mil, 0,3 ml va 0,3 ml). Sd dung que cay vo trong (6.5) dan déu dung dich mu thi trén bé mat thach. 8 yén dia trén mat phdng ngang cho dén khi mu h4p thy hoan toan (khoang 10 min cho dia viva khé). Néu dung dich mau thi khéng hap thyTCVN 11039-6:2015 hoan toan thi G&t Ga trong td Am trong 1 h, np dia huéng lén trén, Sau d6, lat ngugc cae dia va U tir 45h dén 48 h & 35 °C trong td Am (6.4). Lwa chon c4c da cé tir 20 dén 200 khuan lac S. aureus dién hinh, tri khi cht c6 cae dia & 46 pha loding thép hon (> khudn‘lac) xuat hién khudn lac S. aureus dién hinh. Cac khudn lac S. aureus 06 hinh trén, nh&n, Idi, Am ust, duréng kinh tir 2 mm 4én 3 mm trén cae Gia moc thura, mau x4m dén den, bao quanh boi mot quang duc va throng 6 mét vong trang bén ngoai; cac khuan lac keo dinh nhu bo hoac gém khi cham kim cy vao. 9.3. Binh lvgng ‘ém va ghi lai sé khudn lac. Néu trén mét dia co nhidu kiéu khudn lac thi dém va ghi lai s6 long cia tung kigu. C6 thé stv dung cdc dia cia 46 pha long thap nhdt chiva it hon 20 khudn lac. Néu cdc dia 66 nhigu hon 200 khudn lac, gdm cac khudn lac S. aureus dién hinh va cdc khudn lac dién hinh khong xuét hign tai cc 49 pha long cao hon thi str dung cdc dia nay 68 dém S. aureus nhyng khong aém cc khudn lac khéng dién hinh. Chon nhidu hon mét khudn lac ea mdi kiéu da dém va thyc hién phép thir coagulase (8.4.1). Cong tét cd khudn lac cé phan tng coagulase dwvong tinh trén ba dia cla cling 6 pha lofing va nhan voi hé $6 pha long. Bao c&o két qua theo s6 S. aureus trén mot gam m&u thi. 9.4 Nh§n biét S. aureus 9.4.1. Phép thir coagulase Chuy&n cdc khuan lac S. aureus nghi ng& vao cdc éng nhd c6 tir 0,2 ml dén 0,3 mi canh thang TSB chtra natri clorua 10 % va natri pyruvat 1% (5.2), nghién cho d&n khi duc déu. Cy mét ving day huydn phir TSB lén éng mdi tréng thach nghiéng thich hgp, vi dy TSA (6.3). U éng huyén phi nudi cdy trong TSB va éng thach nghiéng 48 cy trong thdi gian tir 18 h dén 24 h & 35 °C. Gilr cde ching cAy trén ba mat nghiéng & nhiét 4 phéng, thye hign cde phép thir lp Iai trong truding hop nghi noe két qua clla phép thir coagulase. Thém 0,5 ml plasma coagulase hoan nguyén véi EDTA (5.6) vao éng huyén phi nuéi cy trong TSB va tron déu. U & 36 °C va kiém tra dinh ki sy déng ty sau mdi 6 h. Phan ung coagulase chi duoc coi la durang tinh Khi dong ty hoan toan va kh6i déng tu khéng di chuyén khi nghiéng hoae ip nguge éng. Trong truéng hyp dng ty mét phan, tude day danh gia la phan ting coagulase dyong tinh 2+ va 3+, cn duge phan tich tiép (xem Tai ligu tham khdo [4)). Thi nghigm ing thoi cdc chiing cdy 44 biét la dong tinh va dm tinh véi cdc chiing c4y nghi ng&, chua r6 vé hoat tinh coagulase, Nhudm Gram tét cd ching edy nghi ng va quan sét hinh thé. Phép thir ngung két latex 66 thé thay thé cho phép thir coagulase néu c&n thir nhanh. 9.4.2 C4c phan tng sinh héa nhan biét bé sung 9.4.2.1 Phan (eng catalase Nhé mét giot hydro peroxit 3 % (5.8) lén phién kinh sach. Lay mét it sinh khéi nuéi ey trong éng thach nghiéng TSA nghién du vao gigt hydro peroxit 3 %. Néu sinh bot khi thi phan ting catalase durong tinh, Phan tng tién hanh ddng thdi vi chting edy 4m tinh va duong tinh. 8TCVN 11039-6:2015 9.4.2.2, Sir dung glucose trong didu kién kj khi Cho vong cdy dy vao day éng canh thang phenol 43 cacbohydrat chira glucose (0,5 %) (5.5). 88 cho chat cdy cham dén d4y éng. Phi ba mat canh thang bang lop dau parafin v6 tring (5.10) voi chidu day it nhat 25 mm. U 5 ngay & 35 °C trong tu Am (6.4). Néu chat chi th di sang mau vang trén toan bé éng thi 6 axit sinh ra trong didu kién kj khi, nghfa la co mat S. aureus. Thyc hién phép thir kiém ching dng thdi (kiém ching ching cy durong tinh, &m tinh va moi trréng). 9.4.2.3 Sir dyng ofmannitol trong didu kign kj khi Tién hanh theo 9.4.2.2, si dung mannitol |am cacohydrat trong mdi tréng. S. aureus thuéng dueng tinh nhung mét s6 ching am tinh. Thye hién phép thir kiém ching déng thai. 9.4.24 Nhay cam voi lysostaphin Dung que cAy I4y khudn lac durgc phan lap tur dia thach vao 0,2 ml dung dich 4m mudi phosphat (5.9) va nghién du. Chuyén mét nia huyén phi vae dng thir hai (13 mm x 100 mm) va tron voi 0,1 ml dung dich dém mu6i phosphat (5.9) am di ching. Thém vao éng ban dau 0,1 ml dung dich lysostaphin (5.7) (64 hda tan trong 100 mi dung dich &ém mudi phosphat 0,02 M chiva natri clorua 1 %-48 06 ndng 46 lysostaphin cudi ciing la 25 pg/ml). U cd hai éng & 35 °C khdng qué 2 h. Néu éng thd nghigm trong thi phép thir durge coi la dvong tinh. Phép thiy 4m tinh néu éng thi nghiém duc. S. aureus thurdng cho két qua dong tinh. 9.4.2.6 Sinh nuclease bén nhigt Khi phan tng coagulase khdng cho két qua ré rang thi cn thy hién phép thir sinh nuclease bén nhiét 46 quan sat sy déi mau tiv xanh lam sang hang tuoi. Day khong phdi la phép thy thay thé cho phan tng coagulase ma la phép thir bd tro, dac bit déi voi cdc phan tng drong tinh 2+, Chuan bj cac lam kinh bang c&ch dan 3 mi thach toluidin xanh-axit deoxyribonucleic (5.6) tran bé mat lam kinh. Khi thach déng dc thi khoan céc giéng duréng kinh 2 mm (tir 10 dén 12 giéng trén mai lam kinh) trén bd mat va loai bé cc nut thach, Nhé khoang 0,01 mi huyén phi vi khuan da gia nhigt (dun séi cach thy 15 min) ‘st dung cho phan tng coagulase vao giéng trén mat thach. U cac lam kinh trong ti 4m 4 h & 35 °C. Néu xuat hién quaing mau héng tuoi réng it nhdt 1 mm bao quanh giéng thi phan tng durong tinh, 9.4.3 Cac dgc tinh Mét sé ac tinh dién hinh cli hai loai Staphylococcus va ¢4c loai Micrococcus durgc néu trong Bang 1.TCVN 11039-6:2015 Bang 1 - Cac dic tinh dién hinh cua S. aureus, S. epidermidis va Micrococcus Bac tinh S. aureus "| S. epidermidis Micrococcus Hoat tinh catalase + + + Sinh coagulase + - - Sinh thermonuclease + + - Nhay cam véi lysostaphin + + 7 Si dung glucose trong didu kién ki khi + + = SG dung mannitol trong diéu kién kj khi + - - (+) Hau nét (@ 60%) cdc ching duzong tinh; (©) Hau hét (@ 90%) cac ching am tinh. 40. Bidu thj két qua Két qua dinh long theo 9.3 dugc bao céo theo sé S. aureus trén mét gam mau thir 14 Bao céo thir nghigm Bao cdo thi nghiém phai chi 15: a) moi théng tin can thiét 48 nhan biét day 44 vé mau they; 'b) phuong phdp l4y mau da sir dung, néu biét; ‘c) phuong phap thir nghiém 44 dung, vién dan tiéu chudn nay, d) moi chi tiét thao tac khéng dugc quy dinh trong tiéu chudn nay hoa nhGng didu dugc coi la tuy chon cing nhu céc sy ob bat ky c6 thé anh hung dén két qua; @) két qua thir nghigm thu durge.TCVN 11039-6:2015 Phy luc A (Quy inh) Thanh phan va chuan bj méi trong va thuéc thir A.A Méitrrong Batrd-Parker A114 Méitreéng co ban Aj44 Thanh phan Trypton 10g Chat chiét thit bd 59 Chat chiét ndm men 19 Natei pyruvat 10g Giycin 12g Liti clorua ngam sdu phan tir nvéc (LICI-6H,0) 59 Thach 209 Nut oft tit A412. Chuan bj Hép dp lc 15 min & 121 °C. pH cuéi clung dat 7,0 + 0,2. Néu can si? dung ngay thi duy tri méi trudng da tan chay & nhiét 66 tt 48 °C dén 50 °C tnvéc khi bd sung tang sinh. Néu khéng can sir dung ngay thi bdo ‘quan méitrudng dong dac 6 4°C + 1 °C dén 1 thang. Lam tan chdy méi trdng truéc khi sir dung. AA.2 Dung dich telurit A424 Thanh phan Kali telurit " (K,TeOs) 41,09 Nuc cdt 100 ml Chi ei dung kali telurit s8n c6 phu hyp di véi phép wir nay (xem A. 1.2.2) A422 Chudn bj Hea tan hoan toan kali telurit trong nude bang cach dun néng rét nhe. Bot kali telurit phai a8 tan. Néu 6 mat chat khong hod tan mau trang trong nuréc thi loai bd bét 4d. Loc qua mang loc c& 15 0,22 um G& khir tring. Dung dich ¢6 thé duoc bao quan ii da m@t thang & nhigt do + 3 °C + 2 °C. Loai 63 dung dich néu cé két tia mau trang.TCVN 11039-6:2015 A143. Dung dich nha tueng long dé trong (ndng 49 khodng 20 % ho&c theo chi din cla nha san xudt), A1.4 Méi trudng hoan chinh AAAA Thanh phan Méi truding oo ban (A.1.1) 100 mi Dung dich kali telurit (A.1.2) 1,0 mi Nhii tuong long 43 trizng (A.1.3) 5,0 ml AA4.2 Chuan bj Lam tan chay méi truéng co ban, sau dé lam ngudi trong ndi cach thuy dén khoding 47 °C. Dui céc didu kién v6 tring, thém dung dich kali telurit va nhG tong long dd trizng, ed hai d& duoc lam 4m trvéc trong ndi cdch thuy & 47 °C, lc ky sau khi thém ting dung dich. AA.5 Chuan bj cc dia thach 'B6 mét lang thich hyp moi tru’ng hoan chinh (A.1.4) vao dia Petri vé trling 8 thu duee moi trong thach day khodng 4 mm va dé cho dac lai, Trréc khi lam khd, cc dia c6 thé durgc bao quan dén 24 h & nhiét 4 + 3 °C + 2°C. CHU THICH: 861 v6i cde dia sn xuat cng nghiép, cAn tuan thé cdc chi dn ciza nha sain xuat vé thei gian baa quan, Trurée hi sir dung, kam kho cac dia, tét nhdt la mé nép ra va up 68 mat thach xuéng dui, dat vao ti Am & nhiét 46 tir 25 °C dén 50 °C, cho dén khi cdc giot nuéc bidn mat trén ba mat cia méi tnzdng. 2 Canh thang trypticase (tryptic) dau twong chia natri clorua 10 % va natri pyruvat 1% A24 Thanh phan Trypticase hoac tryptose 179 Phyton pepton 39 Nati clorua 1009 Kali hydro phosphat (K:HPO,) 259 Dextrose 2.59 Natri pyruvat 109 Nuée edt Ait A2.2 Chuan bj Chinh dén pH 7,3. Dun nhe néu can. Phan phéi 10 mi vao cdc éng nghiém 16 mm x 150 mm. Hap 4p lye 15 min & 121 °C. pH cubi cling dat 7,3 + 0,2, Bdo quan trong mot thang 6 4 °C +1 °C. 12TCVN 11039-6:2015 A3 Thach trypticase (tryptic) dau twong A34 Thanh phan Trypticase pepton 159 Phyton pepton 89 \Natri clorua 59 Thach 159 Nude cat 1 lit A3.2 Chudn bj Gia nhiét 06 khudy d& hda tan thach, Bun séi 1 min, Phan phéi vao cdc dng hoae binh thich hop. Hap Ap lc 15 min & 121 °C. pH cudi cing dat 7,3 + 0,2. A4 Thach toluidin xanh-ADN A441 Thanh phan ‘Axit deoxyribonucleic (ADN) 039 Thech 109 Cansi clorua (dang khan) 4,1mg Nati clorua 10g Toluidin xanh O 83mg Tris(hydroxymethyl)aminometan 619 Nuge ct Ait A42. Chuan bj Hoa tan tris(hydroxymethy!)aminometan trong 1 lit nuéc ct. Chinh pH dén 9,0. Bd sung cdc thanh phan cén lai, triv toluidin xanh © va dun dén séi dé hoa tan. Héa tan toluidin xanh O vao méi truéng. Phan phéi vao cdc binh nén cé nut cao su. Néu sir dung ngay thi khOng can khir tring, Méi trrdng vé tring b&n & nhiét 46 phdng trong 4 thang. AS Canh thang phenol 46 cacbohydrat ASA Thanh phan Trypticase hdc proteose pepton No. 3 109 Netri clorua 59 Chat chiét thit bd (tuy chon) 19 13TCVN 11039-6:2015 Phenol dé (7,2, ml dung dich phenol 43 0,25%) 0,018g Glucose 59 Nude cit 4 iit A5.2. Chudn bj Hea tan glucose vao canh thang co ban nay. Phan phéi c&c phan 2,5 ml vao cde éng nghiém 13 mm x 100 mm cha cdc ng lén men 6 mm x 50 mm dé lat nguge. Hdp dp Ive 10 min & 118 °C. pH cudi cling dat 7,4 + 0,2. Cach khéc, héa tan cdc thanh phan tran, trir glucose, trong 800 mi nude cdt cé dun va thinh thoang khudy. Phan phéi cdc phan 2,0 ml vao cac éng nghiém 13 mm x 100 mm chira céc dng len men 43 lat ngugc. Hap ap lyre 15 min & 118 °C va dé ngudi. Héa tan glucose trong 200 mi nuréc c&t va khiy tring bang cach cho dung dich qua bé loc gir vi khudn. Thém vé tring 0,5 ml dich loc vao mai ‘éng canh thang 64 khir tring sau khi lam ngudi dén 45 °C. Lae nhe dé tron. pH cubi cing dat 7,4 + 0,2 AS Dung dich lysostaphin Hoa tan 2,5 mg lysostaphin trong dung dich mudi dm phosphat 0,02 M chiza natri clorua 1%. A.7 Thuéc thir hydro peroxit Dung dich chira khoang 2,5 g dén 3,5 g hydro peroxit (H0-) trong mi 100 ml. Dung dich nay cé thé chiva cdc cht bdo quan thich hyp nhung ting ndng 46 khéng qué 0,05 %. 14TCVN 11039-6:2015 Th mye tai ligu tham khao [1] TCVN 4830-1:2005 (ISO 6888-1:1999, With Amd, 1:2003), Vi sinh vét trong thc phém vé thie én chan nuéi - Phuong phép dinh lueng Staphylococci cd phan dng duvng tinh coagulase (Staphylococcus aureus va céc foal khéc) trén dia thach — Phan 1: Ky thudt st» dung m6i tring thech Baird-Parker [2] TCVN 4830-2:2005 (ISO 6888-2:1999, With Amd. 1:2003), Vi sinh vat trong thuc phdm va thirs én chan nu6i - Phuong phép dinh Iueng Staphylococci cé phén Ung duong tinh coagulase (Staphylococcus aureus va cdc loai khéc) trén dia thach -— Phan 2: Ky thuét sir dung méi trréng thach fibrinogen huyét tuong thé [3] AOAC 987.09, Staphylococcus aureus in Foods. Most Probable Number Method for Isolation and Enumeration [4] Sperber, W.H. and Tatini, S.R. 1975. Interpretation of the tube coagulase test for identification of ‘Staphylococcus aureus. Appl. Microbiol. 29:502-505 15