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HNG DN PHA MI TRNG NUI CY VI SINH VT

BIN SON XEM XT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

PH DUYT

L L THU TRANG

NG TH M CM

HONG TH MINH HIU

I.

Yu cu:

Ngi thc hin phi nm r MSDS (Material Safety Data Sheet) ca tng
mi trng.

Thao tc phi chnh xc, cn thn, c k nng c bn v thc hnh trong


phng th nghim ha.

V trnh : ti thiu phi tt nghip Cao ng thc phm tr ln. II.


Ti liu lin quan: hng dn cch pha trn nhn sn phm Yu cu an ton Yu cu mi trng III. Thc hnh Cch pha HD1: Hng dn pha mi trng (Nutrien Agar) Mi trng s dng: mi trng Nutrien Agar tng hp. B1: Cn chnh xc khi lng mi trng cn pha cho vo chai duran. pha. B2: Thm nc ct th tch dung dch cn

- eo khu

B3: y kn np chai duran v em tit trng 0 121 C/15 pht, pH cui= 70.2 (250C)

trang, gng tay v o blouse khi - Thao tc nh nhng, hn thao tc. ch pht sinh - B4: Mi trng sau tit - Khng phn rc thi: cc trng ngui n 500C, sau phi mi trng ng nghim rt vo cc ng nghim: vo ng nghim b; mi khi nhit mi trng ri + Th tch mi trng khng trng >500C vi, . qu 1/3 dung tch ng trnh b nghim. bng. + Nghing cc ng nghim sao cho mi trng cch ng nghim ti thiu 3cm.

HD2: Hng dn pha mi trng LSB (Lauryl Sulfate Broth) Mi trng s dng: LSB tng hp B1: Cn chnh xc lng mi trng cn pha cho vo cc thy tinh. B2: Thm nc ct th tch dung dch cn pha, khuy tan. eo khu trang, gng tay v o blouse khi thao tc Thao tc nh nhng khng lm pht sinh bi v rc thi

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HNG DN PHA MI TRNG NUI CY VI SINH VT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

Cch pha

Yu cu an ton

Yu cu mi trng

B3: Phn phi dung dch mi trng vo cc ng nghim c sn ng duham p ngc, mi ng nghim 5ml, vn np ng nghim kn.

eo khu trang, gng tay v o blouse khi thao tc

B4: em cc ng nghim tit trng 1210C/15 pht, pH cui= 6.80.2 (250C) HD3: Hng dn pha mi trng BGBL (Brilliant Green Lactose Bile Broth) Mi trng s dng: mi trng BGBL tng hp. B1: ging HD2, B1

B2: Ging HD2, B2

eo khu trang, gng tay v o blouse khi thao tc

Thao tc nh nhng khng lm pht sinh bi v rc thi

B3: Ging HD2, B3

B4: em cc ng nghim tit trng 1210C/15 pht, pH cui= 7.20.2 (250C)

Tun th ng quy nh trong hng dn vn hnh ni p sut.

HD4: Hng dn pha mi trng EC Broth (EC) Mi trng s dng: mi trng EC Broth tng hp.

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HNG DN PHA MI TRNG NUI CY VI SINH VT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

Cch pha

Yu cu an ton eo khu trang, gng tay v o blouse khi thao tc


0

Yu cu mi trng

B1: ging HD2, B1 B2: Ging HD2, B2 B3: Ging HD2, B3

B4: em cc ng nghim tit trng 121 C/15 pht, pH cui=6.90.2 (250C)

Tun th ng quy nh trong hng dn vn hnh ni p sut.

Thao tc nh nhng khng lm pht sinh bi v rc thi

HD5: Hng dn pha mi trng EMB agar Mi trng s dng: L-EMB Agar B1: ging HD1, B1 B2: Ging HD1, B2 B3: Ging HD1, B3 eo khu trang, gng tay v o blouse khi thao tc

Khng phn
phi mi trng vo a khi nhit mi trng >500C trnh b bng.

B4: Mi trng sau tit trng ngui n 500C, sau phn phi vo cc a Petri v trng, mi a khong 15ml, ng t nhin.

Thao tc nh nhng khng lm pht sinh bi v rc thi

HD6: Hng dn pha mi trng Tryptone Broth (TB) Mi trng s dng: Mi trng tryptone broth tng hp B1: ging HD1, B1 B2: Ging ging HD1, B2 eo khu trang, gng tay v o blouse khi thao tc Tun th ng quy nh trong hng dn vn hnh ni p sut.

B3: Khuy tan mi trng v phn phi 5ml vo mi ng nghim, vn kn np ng nghim. B4: em cc ng nghim hp tit trng 1210C/15 pht.

Thao tc nh nhng khng lm pht sinh bi v rc thi

HD7: Hng dn pha mi trng MR-VP Mi trng s dng: Mi trng MR-VP tng hp Tng t HD6 HD8: Hng dn pha mi trng Simmon Citrate Agar (SCA) Mi trng s dng: Mi trng SCA tng hp Tng t HD1 pH cui= 6.60.2 (250C) Tng t HD1

HD9: Hng dn pha mi trng Tryptose Sulfite Cycloserine (TSC) Mi trng s dng: Mi trng TSC tng hp Page 3/9

HNG DN PHA MI TRNG NUI CY VI SINH VT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

Cch pha B1: ging HD1, B1 B2: Ging HD1, B2

Yu cu an ton

Yu cu mi trng

B3: Hp kh trng 1210C/15 pht, ngui n Nhit pht sinh khong 800C. pH t ni p sut cui=7.60.2 (250C). rt ln c th gy bng

Tun th ng quy nh trong hng dn vn hnh ni p sut.

Kim sot s pht sinh nhit v ting n khi x hi.

HD 10: Hng dn pha mi trng Chapman (CA) Mi trng s dng: Mi trng Chapman tng hp B1: ging HD1, B1 B2: Ging HD1, B2 Tun th ng quy nh trong hng dn vn hnh ni p sut. Nhit pht sinh t ni p sut rt ln c th gy bng

B3: Hp kh trng 1210C/15 pht. pH cui=7.00.2 (250C).

Kim sot s pht sinh nhit v ting n khi x hi.

HD11:Hng dn pha mi trng Trypticase Soy Polymycine B sulfate (TSP) Mi trng s dng: Mi trng TSB tng hp v Polymycine sulfate B B1: ging HD1, B1 B2: Ging HD1, B2 Tun th ng quy nh trong hng dn vn hnh ni p sut. eo khu trang, gng tay v o blouse khi thao tc Khng c ht pipet bng ming.

B3: em hp tit trng 1210C/15 pht, ngui (ddA).

B4:Cn chnh xc 15mg polymycine nguyn cht ha tan vo 10 ml nc ct v trng ta c dd polymycine 0.15% (ddB) B5:B sung ddB vo ddA theo t l 90ml ddA: 1ml ddB, lc u. B6: Dng pipet v trng phn phi mi trng hon chnh pha B5 vo ng nghim v trng c ng duham, 5ml cho mi ng nghim.

Kim sot cc hot ng, thao tc nhm hn ch lng pht thi ra mi trng

HD12: Hng dn pha mi trng thch MYP hon chnh Mi trng s dng: Mi trng MYP tng hp v dd lng trng, polymycine Page 4/9

HNG DN PHA MI TRNG NUI CY VI SINH VT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

Cch pha 0.15%. B1: ging HD1, B1 B2: Ging HD1, B2

Yu cu an ton

Yu cu mi trng

B3: em hp tit trng 1210C/15 pht, ngui n khong 45-500C (mi trng c bn).

B4: Pha mi trng thch MYP hon chnh: b sung vo mi trng chun b B3 theo t l sau: 90ml mi trng c bn: 10ml dd lng trng: 1ml dd polymycine 0.15%, lc u v phn phi 15ml MYP hon chnh vo mi a petri.

Tun th ng quy nh trong hng dn vn hnh ni p sut.

HD13: Hng dn pha mi trng xc nh nm men nm mc v tng s vi khun hiu kh (PDA v PCA) Mi trng s dng: Mi trng PDA v PCA tng hp B1: ging HD1, B1 B2: Ging HD1, B2 Tun th ng quy nh trong hng dn vn hnh ni p sut. B3: em hp tit trng 1210C/15 pht. Lm ngui mi trng n 45-470C trc khi s dng. Nhit pht sinh ra l rt nng do khi thao tc phi m bo tuyt i an ton

Kim sot cc hot ng, thao tc nhm hn ch lng pht thi ra mi trng

HD14: Hng dn pha mi trng Rappaport (RV) Mi trng s dng: Mi trng RV tng hp B1: ging HD2, B1 B2: Ging HD2, B2

B3: Phn phi 10ml mi trng chun b bc 2 vo cc ng nghim v em hp kh trng 1210C/15 pht.

HD15: Hng dn pha mi trng Nitrate Broth (NB) Mi trng s dng: Mi trng NB tng hp B1: ging HD2, B1 B2: Ging HD2, B2

B3: Phn phi vo mi ng nghim 3ml mi trng chun b B2 ri hp kh trng 1210C/15 pht.

HD16: Hng dn pha mi trng Violet Red Bile Agar (VRB) v D-glucose Violet Page 5/9

HNG DN PHA MI TRNG NUI CY VI SINH VT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

Cch pha Red Bile Agar (VRBD)

Yu cu an ton

Yu cu mi trng

Mi trng s dng: Mi trng VRB v VRBD tng hp B1: ging HD1, B1 B2: Ging HD1, B2 B3: B c t vo mi trng pha B2 v s dng my khuy t km gia nhit 1000C trong khong 60 pht lm mi trng tan hon ton. Lm ngui mi trng n 45-470C trc khi s dng. Khng nc trong ni cn kh.

HD17: Hng dn pha mi trng Tryptic Soy Agar (VRB) Mi trng s dng: Mi trng TSA tng hp B1: ging HD1, B1 B2: Ging HD1, B2

B3: Hp tit trng 1210C/15 pht. Lm ngui mi trng n 45-470C trc khi s dng.

HD18: Hng dn pha mi trng KIA Mi trng s dng: Mi trng KIA tng hp B1: ging HD1, B1 B2: Ging HD1, B2

B3: Hp tit trng 1210C/15 pht, pH cui 7.40.2 (250C).

B4: Ch mi trng ngui n khong 500C, phn phi vo mi ng nghim 15ml, sau t ng nghim nm nghing sao cho to c 2 vng trong mi ng nghim: thch nghing nm trn v di l thch ng. ng t nhin hoc nc lnh trc khi s dng. HD19: Hng dn pha mi trng nc peptone m Mi trng s dng: Peptone water tng hp

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HNG DN PHA MI TRNG NUI CY VI SINH VT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

Cch pha pha. B1: Cn chnh xc khi lng mi trng cn B2: Thm nc ct th tch cn pha.

Yu cu an ton

Yu cu mi trng

B3: Phn phi vo cc chai duran hoc ng nghim ty mc ch s dng ri em tit trng 1210C/15 pht.

HD 20: Hng dn pha nc mui sinh l 0.85% Ha cht s dng: NaCl tinh khit Cn chnh xc lng mui tng ng vi th tch dung dch mui cn pha (8.5g mui trong 1 lt nc ct). Khuy tan hon ton v phn phi vo chai duran hay ng nghim ty mc ch s dng.

em hp tit trng 1210C/15 pht.

HD 21: Hng dn pha mi trng Simmon Citrate (SC) Mi trng s dng: Mi trng SC tng hp Tng t HD1 HD 22: Hng dn pha mi trng Tryptone Broth (TB), MR-VP Mi trng s dng: Mi trng TB, MR-VP tng hp B1: ging HD2, B1 B2: Ging HD2, B2

B3: Phn phi mi trng vo cc ng nghim, mi ng nghim 5ml. Sau hp kh trng 1210C/15 pht.

HD 23: Hng dn pha mi trng thch tyrosine Mi trng s dng: thch NA tng hp, tyrosine B1: Cn 0.5g tyrosine v 2g mi trng NA cho vo chai duran.

Thm nc ct cho 100ml. Sau hp kh trng 1210C/15 pht. Cui cng phn phi vo mi ng nghim 10ml mi trng kh trng, nghing ng nghim to thch nghing. HD 24: Hng dn pha mi trng canh thang lysozyme Mi trng s dng: mi trng peptone water, lysozyme

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HNG DN PHA MI TRNG NUI CY VI SINH VT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

Cch pha B1: ging HD2, B1 B2: Ging HD2, B2

Yu cu an ton

Yu cu mi trng

B3: Cn chnh xc 8g peptone water ha tan vo 1 lt nc ct, sau hp tit trng 1210C/15 pht (mi trng 1)

B4: Ha tan 0.1g lysozyme trong 100ml nc ct v trng c HCl 0.01, un si trong 20 pht (mi trng 2). B sung 1% mi trng 2 vo mi trng 1, lc u ri phn phi vo cc ng ng nghim v trng, mi ng nghim 3ml. HD 25: Hng dn pha mi trng Phenol Red Manitol Broth Mi trng s dng: mi trng Phenol Red Manitol Broth tng hp sn B1: ging HD2, B1 B2: Ging HD2, B2 B3: ging HD2, B3

B4: Hp tit trng 1210C/15 pht, pH cui = 7.40.2 (250C). HD 26: Hng dn pha mi trng Ornithine Decarboxylase (ODC) & Lysine Decarboxylase (LDC) Mi trng s dng: mi trng ODC & LDC tng hp sn B1: ging HD2, B1 B2: Ging HD2, B2 B3: ging HD2, B3

B4: Hp tit trng 1210C/15 pht, pH cui = 6.80.2 (250C).

HD 27: Hng dn pha mi trng Glucose OF medium Mi trng s dng: Phenol Red Broth Base tng hp sn

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HNG DN PHA MI TRNG NUI CY VI SINH VT

S hiu: HD-KTT-01 Ln ban hnh: Ngy ban hnh: 02/10/2011

Cch pha

Yu cu an ton

Yu cu mi trng

B1: Pha mi trng Phenol Red Broth Base, hp kh trng 1210C/15 pht, pH cui= 7.40.2 (250C). Lu : ch s dng lng nc ct t hn lng dung dch cn pha 10ml, v 10ml s c dng pha glucose B2 di y.

B2: Ha tan lng glucose cn pha (m bo nng glucose trong mi trng glucose OF lun bng 1%) vo 10ml nc ct v trng lm ngui n nhit 600C, sau un cch thy 600C trong 10 pht. Ha tan 2 dung dch pha B1 v B2, lc u sau phn phi vo cc ng nghim v trng c ng duham p ngc, mi ng 5ml.

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