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Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

LI M U

Vi tnh hnh cc loi bnh dch ang din ra ht sc phc tp. Song song , vn v sc khe cng l mt vn mang tnh thi s nng hi. Chnh v th m khng t ngi t cu hi: Liu trong thi i pht trin nh hin nay th sc khe ca con ngi c c m bo hay khng. Nguyn nhn dch bnh th c rt nhiu nguyn nhn nh do: ha cht, mi trng sng, thc phm cha sn mt s cht c, Nhng quan trng hn ht vn l t vi sinh vt, trong c Staphylococus aureus mt trong nhng nguyn nhn chnh. iu ng lu v quan tm y l chng ny c kh nng tit ra mt s c t bn vi nhit v kh b phn hy nhit cao. Hn na, chng li c kh nng khng methiciline, penicilline khi gp iu kin thun li cn c th ly lan v gy nn nhng cn bnh nguy him. Nhm mc ch tm hiu ngun gc pht sinh, nhng tc hi m Staphylococus aureus gy ra cng nh cc bin php phng nga v cha tr... c bit l tm hiu cc phng php phn tch nhn bit v pht hin chng, em chn ti bo co thc tp Phn Lp Staphylococus Aureus Trn Mu Bnh Nhn.

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Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Phn 1:

TNG QUAN

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Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

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Bi Thu Hoch

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CHNG I: I NT V BNH VIN A KHOA KHU VC HNG NG 1.1. Lch s hnh thnh bnh vin

Hnh 1.1: Bnh Vin a Khoa Khu Vc Hng Ng Cng vi s pht trin nhanh chng ca Th X Hng Ng - ni c th c coi l vng su vng xa, bn cnh Th X Hng Ng li l khu vc bin gii gip vi cc tnh An Giang, Long An v nc bn Campuchia. Bnh vin a Khoa Khu Vc Hng Ng l mt c s trc thuc s Y t ng Thp, chu trch nhim ph trch khm cha bnh 3 huyn: Tam Nng, Tn Hng v Hng Ng. Nhn thy c tim nng pht trin trong tng lai ca Th x cng nh to mi giao ho v Chnh tr - Thng mi gia ngi dn hai nc Vit-Campuchia, y Ban Nhn Dn Tnh ng Thp quyt nh thnh lp Bnh Vin Hng Ng trc thuc S Y T ng Thp trn c s Bnh Vin huyn Hng Ng (c) theo s 16/Q.TL vo ngy 10 thng 3 nm 1994. Bnh vin Hng Ng l bnh vin khu vc ca tnh t ti Th X Hng Ng, tnh ng Thp. Theo tng nhim k bnh vin c s thay i ban gim c v c cu t chc, cn b cng chc cng dn tng theo. Giai on 1994 1995 gim c l Bc s L Hong Cm vi 55 cn b cng chc gm:

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Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm 8 bc s. 11 y s. 11 y t. 5 dc s trung hc. 1 c nhn gy m. 5 n h sinh. 4 k thut vin xt nghim-x quang. 2 k ton trung hc. 1 ti x. 1 k s thit b in nc.

Giai on ban u vi 55 ging bnh, bnh vin c cc khoa nh: Hi Sc cp cu, Ni, Ngoi, Sn, Nhi, Cn Lm Sng, Dc, T Chc Hnh Chnh, Ti chnh k ton. Giai on 1995-1997: gim c l Bc s Phm Vn Kha. Giai on 1998-2000: gim c l Bc s Nguyn Trng Chinh. Giai on 2000 n nay gim c l Bc s Nguyn Minh u. n hin nay bnh vin c m rng vi 233 cn b cng vin chc, bao gm: Tng s CBCNV: Trong : Trn i Hc: i Hc: Trung Hc: S hc: Nhn vin khc: Chi tit (s liu 2010): 2 Thc s. 9 chuyn khoa 1. 27 Bc s. 3 Dc s. 1 iu dng. 10 i hc khc. 15 Y s trung hc. 8 K Thut vin trung hc. 14 Dc s trung hc. 66 Y t, iu dng trung hc. 21 N h sinh trung hc. 5 Trung hc khc. 220/119 n. 12/2 n. 39/15 n. 134/83 n. 11/5 n. 25/14 n.

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Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm 9 iu dng s hc. 3 Dc t s hc. 41 Cn b khc. 28 Hp ng trong qu. C cu t chc: gm 14 khoa phng, trong c 04 phng chc nng, 08 khoa lm sng, 02 khoa cn lm sng. Nhn s: hin c 244 cng nhn vin chc, trong c 141 n.

1.2.

Ban lnh o Gim c: Bc s Nguyn Minh u in thoi: 067.3838423 0913967611 Cc ph gim c: Bc s Quch Trung Ngha in thoi: 067.3837947 0919788144 Bc s V Vn Him in thoi: 067.3838419 0913843963.

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Bi Thu Hoch

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Bi Thu Hoch 1.3.

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Khoa xt nghim 1.3.1. Cn b cng nhn vin ti khoa Chn on Hnh nh Xt Nghim Trng khoa Bs. Trn Quang Vinh. Ph khoa Bs. Phan Thanh Vn. KTV trng khoa Ys. on Minh Vn Trinh Bs. Nguyn Vn To. C nhn Phan nh Huy. C nhn Nguyn Vn Tm. KTV Nguyn Thanh V. KTV Dng Vn Bnh. KTV L Vn Tng. KTV Lm Vn Xun. KTV Thi Ch Linh. KTV ng Th Loan. Ys. L Vn Nhiu H l Ngon Th Thy 1.3.2. Chc nng v nhim v Thc hin cc xt nghim phc v yu cu khi cc khoa ngh. Thng nht p dng thng quy k thut xt nghim theo quy nh, ph bin k thut v kim tra nh k cc thit b. Trin khai thc hin cc dch v xt nghim trong lnh vc y t d phng theo quy nh ca php lut.

Phng Ha Sinh: Tin hnh phn tch cc ch tiu Ha L trong mu theo cc phng php quy nh ca B Y t nh xt nghim nc tiu, huyt hc, sinh ha, HIV, vim gan B Phng Vi Sinh: tin hnh cc bc phn lp, nh danh, lm khng sinh vi cc mu nhn c nh mu phn, mu, m 1.3.3. Cc thit b Chn on Hnh nh Xt Nghim Bng 1: Thng k thit b ti khoa Chn on Hnh nh Xt Nghim My xt nghim huyt t ng T ht v trng My ra film t ng Knh hin vi 2 mt My X-quang cao tng 01 ci 01 ci 01 ci 03 ci 01 ci

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Bi Thu Hoch

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My X-quang ton sng 300mA My siu m trng en My in siu m My sy kh My ly tm nc tiu My phn tch nc tiu t ng My lc T m 37oC, 60 lt T m T sy film T sy 250oC, 120 lt T tr mu 167 lt T tr mu 120 lt Quy lnh My Ion Na+, K+, CLMy ly tm My ly tm a nng 4000 vng/pht My XN sinh ha t ng

01 ci 02 ci 02 ci 02 ci 01 ci 01 ci 01 ci 01 ci 01 ci 01 ci 01 ci 01 ci 01 ci 01 ci 01 ci 02 ci 02 ci 02 ci

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Bi Thu Hoch

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Knh hin vi in t My Ion I

My Ion II T tr mu

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Bi Thu Hoch

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My sinh ha t ng

Hnh 1.2: Mt s hnh nh cc thit b phng Xt Nghim

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Bi Thu Hoch

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Que ly Phn, Dch

T m Que xt nghim tit trng

ng thu mu

Pipet v trng

Hnh 1.3: Mt s hnh nh cc thit b v dng c phng Vi Sinh

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Bi Thu Hoch

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CHNG II: TNG QUAN V VI KHUN STAPHYLOCOCCUS AUREUS 2.1. T cu vng Staphylococcus aureus 2.1.1. Lch s pht hin Ngy 9 thng 4 nm 1881, bc s ngi Scotland Alexander Ogston trnh by ti hi ngh ln th 9 Hi phu thut c mt bo co khoa hc trong ng s dng khi nim t cu khun (Staphylococcus), trnh by tng i y vai tr ca vi khun ny trong cc bnh l sinh m trong lm sng. Staphylococcus aureus do Robert Koch (1843-1910) pht hin vo nm 1878, phn lp t m ung nht v Loius Pasteur (1880) u nghin cu t cu khun t thi k u ca lch s ngnh vi sinh vt (VSV) hc. Nm 1926, Julius von Daranyi l ngi u tin pht hin mi tng quan gia s hin din ca hot ng men coagulase huyt tng ca vi khun vi kh nng gy bnh ca n. Tuy nhin mi n nm 1948, pht hin ny mi c chp nhn rng ri 2.1.2. c im phn loi Bng 2: Bng phn loi khoa hc Staphylococcus aureus Phn loi khoa hc Gii Ngnh Lp B H Ging Loi Eubacteria Firmicutes Cocci Bacillales Staphylococcaceae Staphylococcus Staphyococcus aureus.

Tn khoa hc: Staphyococcus aureus Rosenbach 1884. Trn phng din gy bnh, t cu khun c chia thnh hai nhm chnh: t cu c coagulase v t cu khng c coagulase. S.aureus gy bnh ng c thc phm l t cu c coagulase. Nh enzym ny m trn mi trng nui cy c mu, vi khun to nn cc khun lc mu vng nn cn c gi l t cu vng. Phn loi t cu da trn khng nguyn: Cc t cu c nhiu loi khng nguyn: protein, polysaccharid, acid teichoic ca vch t bo vi khun. Nhng da vo khng nguyn, vic nh loi vi khun rt kh khn.

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Bi Thu Hoch

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Phn loi t cu da trn phage (phage type): t cu c phn vo cc nhm I, II, III, IV. y l phng php s dng nhiu trong phn loi S. aureus. 2.1.3. c im sinh ha Vi khun pht trin d dng mi trng thng thng, hiu kh hoc k kh ty , mc tt 37oC, nhng li to sc t tt 20oC. canh thang sau 5 -6 gi lm c mi trng, sau 24 gi lm c r. mi trng c, khun lc trn, li, bng lng, ng nh, c th c mu vng m, mu vng cam hoc mu trng, tng i ln sau 24 gi. Nhng chng khc nhau lm tan mu nhng mc khc nhau, thch mu typ tan mu thng c quan st xung quanh khun lc.

c tnh sinh ha Catalase Oxidase SAUltex (+) (-) (+)

2.1.4. c im vi khun hc 2.1.4.1. Hnh dng v kch thc

Hnh 1.4: Hnh nh cc vi khun S. aureus di knh hin vi in t. T cu (Staphylococcus bt ngun t ting Hy lp, vi staphyle c ngha chm nho) l nhng cu khun c ng knh khong 1 m, khng di ng v sp xp theo mi hng to thnh cm (t) trng ging nh chm nho. Chng l cc cu khun Gram dng, khng c lng, khng to nha bo v thng khng c v.

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Bi Thu Hoch

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Ngoi ra, cu khun S. aureus khng c kh nng to bo t nh cc vi khun Chlamydomonas perfringens, Chlamydomonas botulinum, v Bacillus cereus cng thng c tm thy trong cc thc phm nhim khun. 2.1.4.2. c t v kh nng gy bnh

Cc loi c t Da theo cc cuc nghin cu v cc bo co khoa hc: T cu vng sn sinh ra 11 c t: c t gy hi chng sc nhim c (TSST-Toxic shock syndrome toxin); c t exfoliatin hay c t epidermolitic; c t alpha; c t bch cu (leucocidin); ngoi c t sinh m (pyrogenic); dung huyt t (hemolysin hay staphylolysin); fribrinolysin (staphylokinase); coagulase; hyaluronidase; lactamase v c t rut (enterotoxin) trong c SEB. C ch gy bnh Ging nh cc vi khun Gram dng khc l Streptococcus v Mycobacteria, S. aureus bm dch vo b mt vt ch nh cc adhensin c bn cht polypeptide. Mt khi bm dnh vo b mt t bo vt ch, tc nhn gy bnh nh S. aureus mi c kh nng khi ng cc qu trnh ha sinh c hiu gy bnh nh tng sinh, bi tit c t, xm nhp v hot ha cc chui tn hiu ca t bo vt ch. S. aureus s tip tc tin su vo trong c th vt ch tip tc chu trnh xm nhp (invasion). S. aureus xm nhp ngoi bo bng cch tit mt s enzym nh: hyaluronidase; hemolysine, leukocidin; exfoliatine .v.v, ph hy cc thnh phn cu to t bo vt ch. Triu chng gy bnh ca Staphylococcus aureus Triu chng ng c thc n do nhim t cu vng Staphylococcus aureus Bnh nhn ng c thc n do t cu xut hin cc triu chng nn i, au qun bng v tiu chy d di, cng v sau phn v cht nn ch yu l nc. Triu chng tiu chy do t cu cng khng km theo mu v t mt nc hn so vi t v E. coli. Bnh nhn khng st hay pht ban, y l c im phn bit gia ng c thc phm do t cu vng vi cc nhm vi khun khc; thn kinh ngi bnh bnh thng. Phn ln trng hp bnh t khi v hi phc trong vng 8-24 gi sau khi pht nhng trng hp nng c th b tt huyt p v gy t vong. Bnh nhn ngoi ra c th b sc do mt nhiu nc v cht in gii. Khc vi ng c thc phm do vi khun thng thng khng gy st hoc st nh, bnh nhn mc ng c do c t SEB ca S. aureus s b st cao. Cc nhim trng da v nim m su L mt hnh thc c bit, nng l inh ru, tip n l chc l, vim ty xng, vim mng phi, vim mng ngoi tim, vim mng no.

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Bi Thu Hoch Nhim trng huyt

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T nhng im nung m, vi khun c th i vo mu v gy nn nhim trng huyt. Nhim trng huyt do t cu l mt bnh thng gp trong bnh vin, thng xy ra nhng ngi c sc khng gim st. Vim rut cp tnh Thng gp cc bnh nhn hay ung thuc khng sinh c khng khun ph rng, khng sinh tiu dit nhng vi khun thng thng rut, lm pht trin chng t cu sinh c t rut v gy nn chng bnh. Hi chng da phng rp (Scalded skin syndrom) Mt s chng t cu vng tit ra c t exfoliatin, gy vim da hoi t v phng rp. Bnh ny thng gp tr mi v tin lng xu. Hi chng shock nhim c (Toxic shock syndrome) Thng gp nhng ph n c kinh nguyt dng bng v sinh dy, nhim bn, b nhim vi khun t cu vng. Bnh kh tr m o v cn nguyn l t cu vng, lin quan n c t gy hi chng shock nhim trng, cy mu khng tm thy t cu vng. 2.1.5. H gen t cu vng Staphylococcus aureus Hin nay ngi ta cng thnh cng trong gii trnh t gen ca cc chng t cu vng c k hiu: Newman, COL, UMRSA 252, MW2, MSSA476, N315, Mu50, RF122 .v.v. [7, 15, 19, 8, 23, 18]. V d: Steven v cs thnh cng trong vic gii trnh t b gen di 2809422bp ca chng S. aureus COL. Kt qu gii trnh t c ghi nhn trn Genbank vi m s: CP000046.1 cho h gen nhn v CP000045 cho h gen plasmid.Theo , trnh t gen ca t cu vng c cha t cc cp G-C, iu ny gy ra mi quan ngi v s chuyn gen t cc chng t cu vng ti cc tc nhn gy bnh Gram dng khc. 2.2. Ni c t rut staphylococcal enterotoxin B. 2.2.1. Cu trc phn t staphylococcal enterotoxin B SEB l 1 trong cc ni c t c sinh ra bi vi khun S. aureus. Thng thng khi b ly nhim vo c th, SEB s tc ng ch yu ln cc h thng vn chuyn ion v nc ca rut, do c gi l enterotoxin (c t rut). c t rut SEB bn vi nhit l tc nhn chnh thng gp nht trong cc v ng c thc phm do S. aureus. c t rut SEB c hnh thnh khi t cu S. aureus sng trong iu kin khc nghit nh nhit mi trng gia tng t ngt, thiu

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Bi Thu Hoch

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2.2.2. C ch gy c ca staphylococcal enterotoxin B Staphylococcal enterotoxin B (SEB) l trung gian kch thch cc lympho T h min dch ca cc vt ch. Cc c t lin kt trc tip n phc hp protein (MHC) lp II trn b mt t bo ch, sau kch thch gia tng s lng ln cc lympho T. SEB c coi l mt "siu khng nguyn ca vi khun v c th to thnh mt cu ni gia MHC lp II ca cc t bo trnh din khng nguyn v vng V ca cc th th t bo T nh CD 4, CD 8; t , kch thch hot ha cc t bo T biu hin cc on gen V m khng cn thit phi c mt qu trnh ch bin v trnh din thng thng. iu ny gy ra s sn sinh mt s lng ln ca cytokine, interleukin 2 (IL2), cc yu t hoi t khi u (TNF-), v cc interferon. Nu n thc phm c SEB bnh nhn c cc triu chng nh chn n, bun nn, nn, v tiu chy. Cc triu chng ny xut hin l do cc cytokine trong cc t bo T ca lng rut c sinh ra t. 2.3. Thc trng nhim Staphylococcus aureus Vit Nam Tnh hnh nhim staphylococcus aureus c mt s s kin: Trong giai on t nhng nm 2008 2009, khi vn V sinh an ton thc phm ngy cng c ch trng, cc cuc nghin cu ca cc c quan TW c y mnh, kt qu l c n hng trm mu thc phm ngy Tt b nhim Staphylococcus aureus trn cc sn phm mc, bnh ko. Vo nm 2010, cuc iu tra v cht lng v sinh an ton thc phm trn cc loi thc phm ng ph nh gi nguy c ly nhim T cu vng l rt cao. Vo nm 2011, hin tng nhim c T cu vng trn thc n lm cho mt b gi Bin Ha (ng Nai) t vong. 2.4. Phng nga, iu tr v x l bnh 2.4.1. Phng nga nhng vng c iu kin v sinh c nhn km, ni ng dn c, c bit l thiu nc sch trong sinh hot, thi tit nng m l iu kin thun li cho bnh pht sinh mnh. - Phng bnh t cu khun cng nh cc bnh nhim khun khc th yu t v sinh c nhn l iu quan trng, ra tay sch trc khi n v sau khi i v sinh l yu cu c bn phng bnh, n ung hp v sinh, nu k trc khi dng, gi gn sch s nhng vt try xc trn da. Nhng bnh nhn tai bin mch mu no nm lit ging c th dn n xut hin nhng vt lot cn phi chm sc h tr vn ng nh xoa

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Bi Thu Hoch

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bp mch mu lu thng, cn i vi bnh nhn i tho ng cn gi gn bn chn trch b phng lot. - Cc bin php nh mang m hoc khn trm u, mang khu trang v ra tay thng xuyn l bin php phng chng ly truyn hu hiu trong bnh vin. - Nhng ngi lm vic trong cc bp n tp th hoc trong cc xng ch bin thc phm cng cn thc hin cc bin php phng h trn. - i vi thc phm th nn bo qun cn thn, khng ln n sng ln vi n chn. Nu nghi ng thc phm b nhim khun th nn loi b. - Khm sc khe nh k cho cng nhn lm vic trc tip vi thc phm, ch khm tai, mi, hng. - Khng nhng ngi b vim xoang, vim mi hng, c mn m tay tham gia ch bin thc phm. - Dng kp khi phc v thc n. - Tng cng ung vitamin, nng cao sc khe, ra tay bng nc nng v x phng, h pH thc phm c ch vi khun pht trin 2.4.2. iu tr Nhiu chng T cu khng vi nhiu khng sinh nht l penicillin nn cn lm khng sinh . C trng hp s dng vaccine bn thn v vaccine tr liu c kt qu. 2.4.3. X l bnh - i vi nhng tn thng khu tr trn da th khng cn phi u tr khng sinh tr khi nhim khun lan rng hoc c bin chng. Ti ch mng m cn lm sch da ti ch bng dung dch st khun, dng khng sinh trc tip bi ngoi da. Cc loi p-xe cn rch m v kt hp dng khng sinh ng ung hoc tim. Nhng loi nhim khun xng, no th cn dng cc loi khng sinh c hiu v khi iu tr th phi theo di cht ch nht l i vi nhng ngi cao tui ngi b x va ng mch hay y tho ng. - Cn s dng nhng loi khng sinh ph hp cho tng bnh nhn. Thng dng gamma globuline iu tr bnh. Mc d S.aureus c th sn xut c men

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Bi Thu Hoch

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penicillinase ph hy vng beta-lactamase lm bt hot cc loi khng sinh nh penicilline G, Ampicilline... Nhng hin nay trong iu tr nhim trng S.aureus ngi ta vn s dng peniclline v cephalosporine khng vi beta-lactamase. - Nafcilline v Oxacilline l hai loi peniclline c dng bng ng tim. Nu nhim trng nng th nn s dng cephalosporine th h th nht nh cephazoline. - Nu bnh nhn khng dung np vi khng sinh nhm beta-lactamase th nn thay bng vancomycine v clidamycine bng ng tim. Dicloxaciline v cephalexine l khng sinh dng ung.

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Bi Thu Hoch

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Phn 2:

VT LIU V PHNG PHP

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Bi Thu Hoch 1.1. Phng php ly v gi bnh phm 1.1.1. Thi gian v a im th nghim:

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Th nghim c lm t u 12/03/2012 n 29/03/2012 ti phng Xt Nghim Khoa Chn on Hnh nh v Xt Nghim Bnh Vin a Khoa Khu Vc Hng Ng - ng Thp. 1.1.2. i tng th nghim Mu mu c ly t ph n ti khoa Sn ca Bnh Vin a Khoa Khu Vc Hng Ng. Trong cng tc phn tch bnh phm chn on mt bnh nghi ng nhim khun; k thut ly; gi bnh phm v k thut nh danh u l nhng giai on quan trng, do cc li lm xy ra bt c giai on no cng u em li kt qu sai lch. Thu thp v vn chuyn bnh phm l mt bc quan trng trong qu trnh phn lp v pht hin cc vi khun gy bnh. Thu thp v vn chuyn bnh phm khng ng cch s nh hng n kt qu chn on cc tc nhn gy bnh v iu tr. Nguyn tc thu thp mu cn bo m: ng chng loi. ng thi gian. ng cch. s lng. m bo cht lng. 1.2. Cch ly bnh phm V tr ly bnh phm V tr ly bnh phm trn bnh nhn c chia lm 2 nhm: a. Ch khng c hn tp vi sinh: gm cc lu cht mu (mu, dch no ty, dch khp, dch mng phi, mng tim, mng bng,), cc mng sinh thit m su. Cn phi kim sot cht ch s ngoi nhim khi ly v phn tch bnh phm. b. Ch c hn tp vi sinh: gm cc cht bi tit (m, phn, nc tiu), nim mc cc l thin nhin, da, phi dng k thut chn lc phn lp vi khun ra khi hn tp vi sinh. Thi gian ly bnh phm Ty thuc vo qu trnh bnh l ca bnh. V kt qu cy khun ch c th hu nghim ng vo thi gian quan trng no ca bnh, trong mt vi trng hp kt qu v nghim t mt mu th duy nht khng hn chn on c bnh nhim khun, m i khi phi ly mu th nhiu ln trong vng 24 48 gi. Phi ly bnh phm trc khi dng thuc khng sinh hay

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Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

sulfamid. Trng hp khng th c, phi ghi r cch tr liu, nu dng penicillin cn thm penicillinase vo mi trng nui cy hay nu dng sulfamides th cn cho thm vo mi trng nui cy cht Paraminobenzoic acid. Qui lut ly bnh phm a. b. c. d. e. f. Khi ly bnh phm cn phi tuyt i tn trng cc quy lut sau: Cc dng c ly bnh phm (kim, ng tim, que bng) phi tuyt i v trng. Cc dng c cha ng bnh phm phi c np y kn v phi v khun ngoi tr l ng phn nhng phi sch. Khng bao gi cho bnh phm chm vo ha cht dit khun no. Tt c bnh phm phi c ghi nhn v phiu th nghim r rng. Phi gi bnh phm ngay n phng th nghim v phi phn tch bnh phm ngay sau khi nhn. Phi cy bnh phm trc khi lm tiu bn hay cc th nghim khc; khi cn phi m t bo trong mt bnh phm lng, phi trn u bnh phm v rt ra mt th tch cn thit trong iu kin tuyt i v khun trc khi quay ly tm bnh phm, sau cn lng ly mu cy v phn nc ni lm cc xt nghim v sinh ha.

Cc phng tin ly bnh phm. Cc phng tin ny ty thuc vo tng loi bnh phm: a. Dng c ly bnh phm: ng tim v kim: ly cc bnh phm cn m xuyn qua da. Dao ly mu: co cc vt au trn da. ng mao qun v khuyn cy khun: ly cht ngoi tit lng bn ngoi. Que bng (2 que): dng ly bnh phm cc l t nhin, nht v. b. Dng c ng bnh phm: Chai c np vn: ng m, nc d dy, mnh m, phn. ng nghim c np vn: ng bnh phm lng. Hp Petri. Chai ln: ng nc tiu tm trc khun lao. 1.3. Cch gi bnh phm Ngoi tr mu, phi cho vo chai hay ng mi trng cy mu ngay sau khi ly. Tt c cc bnh phm khc nn cho vo mi trng chuyn ch Cary Blair vi que bng tm bnh phm. Nu cn gi i xa nn hn np chai mi trng chuyn ch Cary Blair bng paraffin v cho vo mt hp c 2 lp, chn bng cn thn cho khi v; phiu th nghim chn vo 2 lp ca hp. Phi gi ngay bnh phm n phng th nghim trong vng 24 gi.

SVTT: Nguyn Thi Trng

[ 22 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm MU LY V KHUN

Mi trng cy mu lng BHI SPS PABA 37oC/24 gi - 10 ngy C vi khun mc Khng c vi khun mc

Cch 3 ngy cy li trn mi trng thioglycollate lng

C vi khun mc

Khng c vi khun mc

Bo co kt qu : khng c vi khun mc sau khi 10 ngy .

Nhum Gram kho st hnh th vi khun

Cy li trn mi trng thch hp. Lm trc nghim sinh ha. Trc nghim huyt thanh ngng kt.

1. 2. 3. 4. 5. 6. 7.

Staphylococci Streptococci Neisseria gonorrhoeae Neisseria meningitidis Bacillus anthracis Pneumococci Yersinia pestis

NH DANH 8. Brucella 9. Haemophillus influenza 10. Pseudomonas 11. Listeria monocytogenes 12. Enterobacteriaceae tr shigella 13. Leptospira 14. Bacteroides v cc vi khun k kh lin h

S 3: Qui trnh phn lp v nh danh cc tc nhn gy gy bnh t mu ngi

SVTT: Nguyn Thi Trng

[ 23 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Khm vi khun trn BA

Nhum gram: Cu khun gram (+)

Catalase (+)

Catalase

Catalase (-)

Oxidase

nh danh theo qui trnh Streptococci Oxidase (-) Cy tng sinh trn BA

Oxidase (+)

Micrococci

Chn khm vi khun mc

COAGULASE STAPHYLATEX COAGULASE (+) COAGULASE (-)

Staphylococcus aureus

Staphylococci coagulase (-) (SCN)

Khng NOUVOBIOCIN SCN khc (-)

S. Saprophyticus (+)

S 4: Qui trnh th nghim sinh ha pht hin Staphylococcus aureus

SVTT: Nguyn Thi Trng

[ 24 ]

Bi Thu Hoch 2.1. Cy mu 2.1.1. Ch nh cy mu

Ngi hng dn: CN. Nguyn Vn Tm

Phi ch nh cy mu trc cc trng hp nhim trng c th c du khun huyt tm thi (transient bacteremia) hay nhim trng huyt (septicemia). Do vy, nn ch nh cy mu trc cc bnh nhn c mt trong cc triu chng nh: st, n lnh, lnh run, ting thi tim (cardiac murmur) nghi ng vim ni tm mc, c xut huyt da hay nim mc, xut huyt dng sao (splinder) trn mng tay, chong. 2.1.2. Thi im cy mu Phi cy mu trc khi bnh nhn dng khng sinh h thng. Trong bnh vin, bc s phi cho cy mu trc khi bt u cho bnh nhn dng thuc khng sinh. Tuy nhin trong cc trng hp bnh nhn ang iu tr khng sinh nhng cc triu chng ca du khun huyt hay nhim trng huyt vn khng thuyn gim th bc s cng nn cho ch nh cy mu pht hin tc nhn vi khun gy nhim trng. Thi im tt nht cy mu l khi bnh nhn b n lnh hay ang lnh run trc khi st, hay lc bnh nhn ang ln cn st. C th cy mu 2 ln trong vng 1 gi u. 2.1.3. Cch ly mu cy Ly mu tnh mch bng phng php v trng (st trng da bng cn 70%, ch kh ri mi chc kim ly mu). Th tch mu c ly cy chim 1/10 th tch mi trng cy mu. Thng thng ly 3-5ml mu cy vo mi trng cy mu c th tch 50ml. i vi trng hp bnh nhn l tr em, ly ch 2-3ml cng c. 2.1.4. Mi trng cy mu BHI, TSB, hay Columbia Broth cho vi khun hiu kh. cy ym kh, thm vo mi trng cc cht kh nh Thioglycollate, L cystein. khng ng, tt nht l dng Sodium Polyanethol Sulfonate (SPS), nu khng c th dng citrate hay heparin. 2.2. Theo di cy mu Chai cy mu c trong t m 35oC hay 37oC v theo di mi ngy trong 7 ngy xem c du hiu vi khun mc hay khng trong mi trng cy mu lng: (1) c ht ng trn mt hng cu, (2) c u hay c mng, (3) tiu huyt, (4) ng huyt tng, (5) c gas, (6) c ht trng trong lp hng cu hay mt lp hng cu.

SVTT: Nguyn Thi Trng

[ 25 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Nu l chai cy mu c 2 phase th trc khi v mi ngy sau khi quan st mt thch ca phase c xem c khm vi khun mc hay khng; nu khng c khm vi khun mc trn phase c, nghing trng phase lng trng ln phase c. Bt c lc no pht hin c du hiu vi khun mc hay nghi ng vi khun mc, tin hnh cy phn lp ngay, tt nht l trn thch b dng nht l thch nu c b sung (CAXV = Chocolae Agar Isovitex), nu khng c th cy trn thch mu BA hay thch nu CA, ng thi lm mt pht nhum Gram kho st trc tip. Nu kt qu nhum Gram thy c vi khun, c th lm khng sinh trc tip t chai cy mu. Nu trn phase c c vi khun mc th tin hnh nh danh v lm khng sinh . C vi khun rt d b ly gii sau khi mc trn mi trng lng nh S.pseumoniae, do nu c th th nn lm mt cy truyn m ln mt thch phn lp sau khi chai cy mu qua m hoc sau 24 gi. Sau 7 ngy theo di chai cy mu, phi cy truyn m mt ln na chc chn trc khi tr li cy mu m tnh. Cc vi khun hay nm thng l cc tc nhn nhim trng huyt hay nhim nm huyt Gram (-) Escherichia Coli Klebsiella spp. Enterobacter spp. Proteus Salmonella typhi Samnella ( ngoi S.typhi) Pseudomonas aeruginosa Neisseria meaningitidis Haemophilus influenza Bacteroides fragilis Brucella spp. Pseudomonas pseudomallei 2.3. Vn vi khun ngoi nhim T da: Staphylococcus epidermidis Propionobacterium acnes Gram (+) v nm men Staphylococcus aureus S.epidermidis S.pyogenes (nhm A) S.agalactiae (nhm B) Streptococci, tiu huyt (viridians) Streptococcus pneumonia E. faecalis (nhm D) Listeria monocytogenes Clostridium perfringens Peptococcus spp. Peptostreptococcus spp. Candida albicans v cc nm men khc

SVTT: Nguyn Thi Trng

[ 26 ]

Bi Thu Hoch Diphtheroides. T mi trng xung quanh: Clostridium spp. Acinetobacter spp. Bacillus spp.

Ngi hng dn: CN. Nguyn Vn Tm

Loi tr c ngoi nhim khi kt qu cy mu cho thy vi khun: Cng phn lp c t 2 chai mu cy t mt bnh nhn. Cng phn lp c t mt bnh phm khc cng trn bnh nhn . Vi khun mc nhanh. Cc dng vi khun c cng type sinh hc v khng nh nhau vi khng sinh. Tr li kt qu cy mu dng tnh Tt c cc vi khun mc c trong chai cy mu u phi tr li cho bc s iu tr. Tham kho vi bc s lm sng rt kinh nghim trong cc trng hp ngoi nhim. Cc vi khun nghi ng l ngoi nhim, khng cn thit phi lm khng sinh v thng bo cho cc bc s bit s nghi ng ny. Bng 3: Nhng vi khun c th tm thy trong m v dch ca c th STT M hay bnh phm Mu Vi khun thng thy Vi khun gy bnh

Khng c (bnh phm c th b nhim bi cc vi khun thng sng trn da)

1. 2. 3. 4. 5. 6. 7.

Staphylococcus aureus Hemolytic streptococci Enterococci Pneumococci Neisseria menigitidis Neisseria gonorrhoeae Cc vi khun ng rut ngoi tr Shigella 8. Cc loi Pseudeomonas 9. Haemophilus influenza 10. Yersinia pestis 11. Brucella 12. Bacteroides v cc vi khun k kh lin h 13. Listeria cynomotogenes 14. Bacillus Anthracis 15. Leptospira 1. Hemolytic Streptococi thng

Mi, yt

1. Hemolytic

SVTT: Nguyn Thi Trng

[ 27 ]

Bi Thu Hoch hu, cung hng v ming

Ngi hng dn: CN. Nguyn Vn Tm Streptococi 2. Neisseria catarrhalis 3. Staphylococci (coagulase m). 4. i khi Staphylococus aureus. 5. Haemophius haemolyticus. 6. Haemophilus 7. influenza 8. Dipthroid bacilli. 9. Non hemolutic Streptococci. 10. Pneumococci 11. i khi Coliform bacilli l nhm A, i khi nhm B v C 2. Corynebacterium diptheriae 3. Bordeltella pertussis 4. Meningococci. 5. Staphylococus aureus (coagulase dng) khi c nhiu. 6. Haemophilus influenza khi c nhiu. 7. Coliform bacilli khi c nhiu. 8. Pneumococci khi c nhiu. 9. i khi Yersinia pestis. 10. Vi khun loi Fusospiochets

Ging nh yt hu, cung hng v ming

1. Pneumococci khi c nhiu 2. Klebsiella Pneumonniae. 3. Hemolytic Streptococci khi c nhiu. 4. Hemolytic Streptococci 5. Staphylococcus aureus khi c nhiu. 6. Yersinia Petis. 7. Mycobacterium tuberculosis 8. Bordeltella pertussis 9. Coliformbacilli cc loi 10. Pseudeomonas cc loi 11. Proteus khi c nhiu 1. Staphylococus aureus (coagulase dng). 2. Staphylococus pyogenes 3. Coliform bacilli 4. Enterococci 5. Cc loi Proteus 6. Cc loi Pseudomonas 7. Cc loi Clostrdium 8. Bacteroides [ 28 ]

Vt thng, Cc loi vi khun thng Nhc bc v thy trn da vt phng 1. Diptheroid bacilli 2. Staphylococci (Coagulase m) 3. Coliform bacilli 4. Cc loi bacilli

SVTT: Nguyn Thi Trng

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm 9. Cocci k kh 10. Corynebacterium diptheriae him thy. 11. Bacillus anthracis him thy

C quan bi tit, c quan sinh dc, niu o v m h.

1. Coagulase m Staphylococci 2. Coliform bacilli 3. Enterococci 4. DorderleinS bacilli 5. Mycobacterium khng gy bnh 6. Haemophius Vaginalis. 7. Bacteroides 1. 2. 3. 4. 5. 6. 7. Escherichia coli Enterococci Bacteroides Clostridia Staphylococci Cc loi proteus Cc loi Pseudomonas 8. Cc loi Klebsiella 9. Cc loi Enterobacter 10. Cc loi Serratia

1. 2. 3. 4.

Neisseria gonorrhoae Treponema pallidum Haemophius ducreyi Haemophius Vaginalis khi c nhiu. 5. Listeria monocytogenes 6. Enterococci khi c nhiu 7. Donovania granulomatis

Phn

1. 2. 3. 4. 5. 6. 7. 8.

Cc loi Samonnela Cc loi Shigella Cc loi Arizona Vibrio cholera Vibri parahemolyticus Escherichia coli Staphylococci khi c nhiu Cc loi proteus khi c nhiu nht l tr em. 9. Cc loi Pseudomonas khi c nhiu nht, nht l tr em. 10. Cc loi Providencia 11. Citrobacter 12. i khi Serratia. 13. i khi Enterobacter. 14. i khi Klebsiella. 15. Edwardsiella tarda 1. Escherichia coli. 2. Klebsiella Enterobacter Serratia. 3. Cc loi Proteus cc loi Providecia. 4. Pseudomonas aeruginosa 5. Enterococci (Streptococus fecalis) khi c nhiu. 6. Coagulase Possitive v negative Staphylococci. 7. Cc loi Haemophius, c th l Haemophius Vaginalis. [ 29 ]

Nc tiu

1. Staphylococci (Coagulase m) 2. Diptherroid bacilli 3. Coliform bacilli 4. Enterococci 5. Cc loi Bacilli 6. v Hemolyticus streptococci. 7. Cc loi Proteus 8. Lactobacilli

SVTT: Nguyn Thi Trng

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm 8. Hemolytic streptococci thng l nhm B v D. 9. Neissinia gonorrhoeae 10. Mycobacterium tuberculosis. 11. Cc loi Leptospira. 12. Cc loi Alcaligenes. 13. Cc loi Samonella.

3.1. Cc th nghim sinh ha nh danh 3.1.1. Th nhim -Lactamase Nguyn tc Nhng vi khun c sinh ra -lactamase s lm a thuc th cha di cht l Nitrocefin, l mt khng sinh thuc h Cephalosporin i mu t vng sang . Kt qu th nghim ny d dng pht hin bng mt thng. Cch thc hin Dng kp sch ly mt a -lactamase qut trc tip ln khun lc ca chng vi khun cn th nghim. Quan st s i mu ca -lactamase. Cch c kt qu: [ - ]: nu a khng c s thay i mu. [ +]: nu qut khm vi khun trn a chuyn sang t mu vng sang mu . hu ht cc vi khun th s thay i mu khng qu 5 pht. Tuy nhin, mt s chng Staphylococci c th n 1 gi.

Vi khun

Kt qu

Thi gian c KQ

Tng ng vi KQ Khng sinh

khng vi Penicillin, Ampicillin, Carbenicillin, Ticarcillin. Staphylococcus aureus [+] 1 gi C th nhy vi Cephalothin, Methicillin, Oxacillin, Nafcillin v cc Penicillin khng Penicillinase. khng vi Ampicillin, nhy vi Cephalosporin.

H. influenza

[+]

1 pht

SVTT: Nguyn Thi Trng

[ 30 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

N.gonorrhoeae M.catarrhalis

[+]

1 pht

Khng vi Penicillin

E.faecalis

[+]

5 pht

Khng vi Penicillin v Ampicillin C l l Bacteroides spp. C th khng vi Penicillin v Cephalosporin k c Cefotaxime v him vi Cefoxitin.

Vi khun ym kh

[+]

30 pht

3.1.2. Trc nghim Catalase: Cng dng: Chn on phn bit khm khun Staphylococci vi khm khun Pneumonocci v Steptococci. Nguyn tc: T cu khun c enzyme catalase s lm phng thch O2 t nc Oxy gi to hin tng si bt. Catalse H2O2 K thut: (trn knh) Nh mt git nc Oxi gi 30% trn mt knh ng vt sch. Chm khuyn cy v khun vo khm vi khun tinh khit mun th, ri cho khuyn chm vo git H2O2. Quan st s si bt ngay tc khc nu l phn ng dng. Kt qu: H2O +
1 2

O2

Ging Staphylococci Pneumonococci Streptococci

Catlase + -

SVTT: Nguyn Thi Trng

[ 31 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Ch : + Trnh khng c khuyn cy chm vo thch mu ly theo hng cu trong thch, v hng cu ny s lm phn ng dng gi. + Khuyn cy khun bng platin s cho phn ng dng gi. 3.1.3. Kh nng tng trng v ln men trn mi trng M.S.A (Chapman) Cc loi T cu khun u tng trng c trn mi trng MSA cha 7,5% NaCl. Ring gc gy bnh Staphylococcus aureus do c kh nng ln men c trn ng Mannitol nn s to mt vng mu bao quang khm vi khun. 3.1.4. Trc nghim Coagulase Cng dng: c dng nh danh Staphylococcus aureus Nguyn tc: Vi khun Staphylococcus aureus c enzyme coagulase c kh nng lm ng huyt tng. Loi coagulase kt dnh c xc nh bng k thut trn knh. Cn loi coagulase t do c xc nh bng k thut trong ng nghim k thut. K thut trong ng cho kt qu chc chn hn k thut trn knh. 3.1.5. Trc nghim Novobiocin: Cng dng: dng nh danh Staphylococcus saprophyticus. Nguyn tc: Staphylococcus saprophyticus khng c c vi Novobiocin. Nu dng a giy tm 5g nouvobiocin (phng php khuch tn khng sinh trn thch) ng knh vng b chn i vi Staphylococcus saprophyticus nh hn 16mm. K thut: Cy vi khun dng staphylococci cn phn loi ln mt thch T.S.A t a giy ta,63 Nouvobiocin ln trn. 37oC trong 18 24 gi. o ng knh vng trng b chn. Ging Stap.aureus Stap.epidermidis Stap.saprophyticus Catlase + + -

SVTT: Nguyn Thi Trng

[ 32 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Bng 4: Phn bit mt s loi Staphylococcus gy bnh S.aureus Mi trng Chapman Tng trng Acid ha Mannitol + Acid (vng) S.epidermidis + S.saprophyticus +

Thay i

Thay i

Mu sc khm khun Nhy cm in viro vi Nouvobiocin

Xm, kem, hoc vng

Trng

Trng

3.1.6. Th nghim SAUTlATEX (STAPHYLATEX) Nguyn tc: Vi khun Staphylococcus aureus phn bit c vi cc vi khun Staphylococci Coagulase [-] khc l c protein A trn vch t bo v c yu t kt cm (clumping factor) trong khi cc vi khun Staphylococci Coagulase [-] khng c cc yu t ny. SAUlatex, thuc th A pht hin c protein A, v thuc th B pht hin c yu t kt cm ca S. aureus. Th nghim da trn nguyn tc ca phn ng t, thc hin c d dng trn lame soi knh hin vi, v kt qu c bng mt thng. Thnh phn thuc th: SAUlatex gm cc thuc th sau: Thuc th A: l dung dch laex dng pht hin protein A trn vch ca vi khun S. aureus m khng gp trn cc vch ca vi khun Staphylococci Coagulase [-] khc. Thuc th B: l dung dch thuc th pht hin cc yu t kt cm c trn S. aureus Hp SAUlatex cha dng hay sau khi dng phi gi trong t lnh 4oC, SAUlatex bn n 2 nm k t sau ngy sn xut. Phng php thc hin th nghim

SVTT: Nguyn Thi Trng

[ 33 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Chun b mt lame soi knh hin vi mi, lau sch bng cn, ri sau lau kh. Nh khi cm lame phi cm trn cnh lame ch khng cm trn mt lame. Dng bt m hay bt acetone k trn lame hai 18 x 18mm k nhau. Nh trn gc 2 , mi mt git nc mui sinh l hay nc ct th tch c 10-20l. Lm mt huyn dch vi khun th nghim tht c u trong git nc mui sinh l hay nc ct ny, trnh c cn. Nh ln mt gc khc ca vung 1 git thuc th A cng c th tch c 10l. Dng vng cy hay que tm g sch trn git huyn dch vi khun vo git thuc th A, trn u trong 30 giy n 1 pht ri quan st bng mt thng xem c hin tng t hay khng. Nu c, vi khun th nghim c xc nh l S. aureus. Nu khng c hin tng t, nh tip vo mt git thuc th B cng c th tch 10l vo th hai, ri cng dng que g sch hay vng cy trn tip trong 30 giy n 1 pht na. Nu c hin tng kt t, vi khun th nghim c xc nh l S. aureus. Nu vn khng kt t, xc nh vi khun th nghim l staphylococci coagulase. 3.2. Phng php nhum Gram 3.2.1. Nguyn tc Khi nhum vi khun vi thuc mu Crystal Violet v mt cht gn mu Iodine, nu ta ty mu bng cn 95%, c mt s vi khun vn gi cht mu tm ca Crystal Violet, l nhm Gram (+). Mt s khc b ty mt mu tm v mng t bo thiu cht chuyn bit dng kt hp vi Crystal Violet, l nhm Gram (-). Khi nhum li vi dung dch Safranin 0,5%, nhm Gram (+) vn gi mu tm, cn nhm Gram (-) s bt mu . 3.2.2. Dng c v thuc nhum: Dng c Khuyn cy khun. n cn. Knh ng vt (lame) sch. Bt ch m. Chai nh git ng nc mui 0,85% ng h pht. Thuc nhum Dung dch Crystal Violet. Dung dch Iodine. Cn 95%. Dung dch Safranin 0,5% hay dung dch Carbon Fuchsin 1/10. 3.2.3. K thut

SVTT: Nguyn Thi Trng

[ 34 ]

Bi Thu Hoch Mu th lm phin pht

Ngi hng dn: CN. Nguyn Vn Tm

Nu l la cy lng: ty theo c ca la cy, ta ly 1 git nh hay ly y 2,3 khuyn cy khun lm phin phit. Nu l la cy trn mi trng c: dng khuyn cy khun chm 1 khm vi khun ring r, lm huyn trc trong 1 git nc mui trn knh, xong tri mng thnh phin pht. Nn phn tn u, khng dy, c th quan st c tng t bo vi khun. K thut lm phin pht. Cm que cy tht thng ng, u khuyn t trn ngn la n cn v t u que cy. ngui ly 1 khuyn cy y bnh phm. t khuyn bnh phm vo gia tm knh, cho p sut vo mt knh, pht u trn knh thnh nhng hnh bu dc vi din tch 1,5 x 2,5cm. t u que cy kh khun. Lu nh phin pht Phin pht phi c kh t nhin trong khng kh, hoc h nh trn ngn n. Sau , lu nh phin pht bng cch a nhanh tm knh qua li trn ngn la 4 5 ln, mt c phin pht pha trn. Khng c h qu nng s lm cc t bo co li. ngui trc khi nhum v khoanh vng bnh phm bng nt ch m d tm. K thut nhum Ph dung dch Crystal Violet ln phim pht, 1 pht. Ra nc v nghing knh cho ro. Ph dung dch Iodine: Ln I 30 giy, b. Ln II 30 giy, b. Ph cn 95% ln phin pht, 30 giy. Ra nc v nghing knh cho ro. Quan st phin pht, nu cn nhng vt mu tim m phi ty mu ln na. Nhum li vi dung dch Safranin trong 30 giy. Ra nc, kh v kho st vi vt knh du. 3.2.4. Kt qu Theo phng php nhum Gram, vi khun c phn bit thnh 2 nhm: Vi khun Gram (+): gi mu tm ca Crystal Violet. Vi khun Gram (-): gi mu ca Safranin. 3.3. K thut lm khng sinh

SVTT: Nguyn Thi Trng

[ 35 ]

Bi Thu Hoch 3.3.1. Nguyn tc

Ngi hng dn: CN. Nguyn Vn Tm

Thuc khng sinh thm trn a, khuch tn ra mi trng xung quanh ngn chn s pht trin ca vi khun. ng knh vng b chn din t tnh cm ng ca vi khun i vi thuc khng sinh, trng hp khng c vng b chn, v khun khng li thuc khng sinh. 3.3.2. Chun b a khng sinh L nhng a giy c ng knh 6mm c tm mt dung dch thuc khng sinh vi nng tiu chun. a khng sinh phi c gi t lnh, c km theo cht lm khan nc, chng m sc nng v s ngoi nhim lm hng a. Mi trng Mi trng tiu chun l MHA (Mueller Hinton Agar) pH: 7,4. Vi cc vi khun kh tnh phi cho thm 5% mu. Mi trng c vo hp Petri vi b dy ca thch l 4mm. Bo qun t lnh v khng c dng sau 7 ngy. Trc khi dng, phi ly ra khi t lnh v cho n khi t c nhit phng. Dung dch chun c. L dung dch mui BaSO4 Dung dch d tr A BaCl2.2H2O ED qsb Dung dch d tr B H2SO4 Dung dch chun c Dung dch A Dung dch B Vi khun th nghim 11,7g 1000ml 1% 0,5ml 99,5ml

Vi khun sau khi c cy thun khit, cy vo ng 2-5ml mi trng TSB (BHI) 37oC trong 2-8 gi t c c chun ca mui Barium Sulfate. 3.3.3. K thut Cc bc thc hin: Chn khm vi khun. Lm huyn dch vi khun trong nc mui hp v khun hay mi trng lng. Sau khi so vi c chun.

SVTT: Nguyn Thi Trng

[ 36 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Dng que gn v trng nhng vo huyn dch vi khun chun b v p kit nc trn thnh ng. Tri u vi khun trn mi trng thch. Thch trn chun thng dng Mucller Hinton (MHA). Dng kp v trng ly cc a khng sinh gn trn mt thch, mi hp t khong 7-8 a. 37oC v c kt qu sau 24 gi. o ng knh vng khun. c kt qu bng cch so vi bng chun. Bng 5: Mt S Tiu Chun Bin Lun ng Knh Vng V Khun V im Gy Nng c Ch Ti Thiu (MIC) Ca Enterrobactorlaceae a Khng Sinh M Hm Lng (Code) ng knh vng vi khun o bng mm trn Khng Trung gian 15-16 14-16 14-16 14-17 Nhy

Amikacin Ampicillin Amoxlcallin Amoxlcallin/clavuanic acid CEPHALOSPORIN v CEPHEM khc Cephalexin Cefamandole Cefepime

Ak Am Ax Ac

30g 10g 10g 20/10g

14 13 13 13

17 17 17 18

Cp Cd Cm

g 30g 30g

14 14

15-17 15-17 15-17

18 18 18

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[ 37 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Cefoperazone Cefuroxime Cefutriaxone Ceftazidime Cefuroxime acetil Cefaclor Imipenem Gentamicin Kanamycin Netilmicin Tobramycin Streptomycin Tetracycline Doxyciline Ciprofloxacin Norfloxacin

Cf Ct Cx Cz Cu Cr Im Ge Kn Ni Tb Sm Te Dx Ci Nr

75g 30g 30g 30g 30g 30g 10g 10g 30g 30g 10g 10g 30g 30g 5g 10g

15 14 13 14 14 14 13 12 13 12 12 11 14 12 15 12

16-20 15-22 14-20 15-17 15-22 15-17 14-15 13-14 14-17 13-14 13-14 12-14 15-18 13-15 16-20 13-16

21 23 21 18 23 18 16 15 18 15 13 15 19 16 21 17

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[ 38 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Ofloxacin Nalidixic acid Trirnethoprim/sulfamethoxazole Vancomycin Chloramphenicol Nitrofurantoin Penicicllin ( dng Oxacllin) Clindamycin Erythromycin

Of Ng Bt Va Cl Fr Ox cL Er

5g 30g 1,25/23,75g 30g 30g 300g 1g 2g 15g

12 13 10

13-15 14-18 11-15

16 19 16 15

12 14 14 13

13-17 15-16 15-20 14-22

18 17 20 21 23

Hnh 2.1: a khng sinh

Hnh 2.2: Mi trng MHA [ 39 ]

SVTT: Nguyn Thi Trng

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

4.1. Cch pha ch mt s mi trng nui cy vi khun Mi trng l nhng thc n nui cy vi khun. Mi trng phong ph i vi vi khun l mi trng gip cho cc vi khun tng trng mnh v nhanh. Mun c nh th, mi trng phi hi cc iu kin sau y: Giu cht dinh dng thch hp: nito, carbon, protid, acid amine, mui khong, yu t tng trng. iu kin vt l thun li: nhit , pH, p lc thm thu. C nhiu cch phn loi mi trng. Theo dinh dng v cch s dng, mi trng c chia lm 6 nhm: 4.1.1. Mi trng dinh dng c bn. Mi trng b. Mi trng chuyn ch. Mi trng phong ph. Mi trng phn lp. Mi trng nh danh hay mi trng sinh ha. Mi trng dinh dng c bn.

iu ch nc tht Nc tht tt, phi trong. Mun t c kt qu ny cn phi c nhng iu kin: Chn tht b t. Lc ht m gn. Tht khng b ng t lnh. Xay tht nh. Cn v thm nc cho ng. t lnh t 17-20 gi.

Dng c Dao, tht, my xay tht, ni ng tht (bng inox, khng r), gi mc tht, nc lc, ng ong nc, vi lc, giy lc, phu. Cch lm 1kg tht b lc ht m, gn, ct ming nh, em xay nhuyn. Cho tht vo ni, ong nc ( lnh mt m) theo t l: Tht xay nhuyn Nc ct Men (nu c) 500g 1000ml 2 ng.

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[ 40 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Khuy u, y np ni. vo t lnh 17-24 gi. Ngy hm sau, ly ra, un t t ln 100oC, si 30 pht. Lng 5 pht, lc. Nn lc qua vi t v 2 lp giy lc trnh mng m trn mt. Trc khi lc, nht t bng thm nc vo l trn ca ng phu giy lc khng b rch. Cho vo chai hp 121oC trong 15 pht. 4.1.1.1. Canh thang L mi trng cn bn ca vi khun hc, mu vng nht. Thnh phn gm c: nc tht, peptone, v NaCl. Nc tht Peptone Martin NaCl 600ml 500ml 0,5g

un si cho tan, iu chnh PH 7,4 bng NaOH 10%, hp 121oC trong 30 pht. Ngy hm sau gn ly nc trong, lc qua giy lc t nhiu ln, phn phi vo ng nghim hay cu bnh, hp 110oC trong 10 pht. 4.1.1.2. Thch thng Nc tht Peptone Martin NaCl Thch 500ml 500ml 0,5g 20g

Cho thch vo hn hp nc tht, pepetone bt, mui, un si cho tan u, chnh pH = 7,4. Cho vo mt bnh thnh thng ng em hp 121oC trong 30 pht. Ngy hm sau ct b phn cn di, ly phn thch trong cn li ct nh, un si, lc qua vi hay bng gn khi cn nng, phn phi vo ng nghim 19x150, 7ml mi ng. Hp 110oC trong 15 pht, ly ra nm nghing. 4.1.2. Mi trng b. Thch mu L mi trng cn bn dng phn lp v nui cy cc vi khun kh tnh. Cao tht b 10g Peptone 10g NaCl 5g Thch si 20g pH = 7,4 Cn cc thnh phn, ha tan vi 1 lt nc ct un si, iu chnh pH = 7, 3. Hp kh khun 121oC trong 15 pht. Ly ra mi trng ngui 50oC, cho thm 5% mu ti (cu, th, ngi). Trn u phn phi ra hp Petri v khun. ngui, lt ngc hp thch ct vo t 37oC. Hm sau ly ra loi b nhng hp thch ngoi nhim, nhng hp cn li ct vo t lnh dng dn.

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[ 41 ]

Bi Thu Hoch Mi trng T.S.A (Trypticase Soy Agar) pht. Mi trng BHI pht. 4.1.3. Mi trng chuyn ch Mi trng Cary Blair Trypticase peptone Phytone peptone NaCl Thch si pH =7,3

Ngi hng dn: CN. Nguyn Vn Tm

15g 5g 5g 15g

Cn 40g bt TSA ha trong 1 lt nc ct, un si cho tan. Hp 121oC trong 15

Dch ty b con (Calf-Brain) Tim b Peptone NaCl Disodium Phosphate (Na2HPO4)

200g 250g 10g 5g 2 ,5g

Ha tan 37g mi trng kh vo 1 lt nc ct. Hp kh khunn121oC trong 15

Dng chuyn ch hu ht cc loi bnh phm. Sodium thioglycollate 1,5g Disodium Phosphate 1,1g NaCl 5g Thch 5g Ha hn hp trn vo 991ml nc ct un si, khuy cho tan. ngui 50oC, cho thm 9ml dung dch calcium chloride 1%, khuy u trong 2 pht. iu chnh pH = 8,4 vi NaOH.N hay HCl.N. Phn phi vo chai c bit, mi chai 7ml. Sau hp 110oC trong 15 pht. 4.1.4. Mi trng phong ph ha Mi trng Tetrathionate lng Dng lm tng sinh s lng vi khun Samonella shigella trong phn. Tetrathionate cn bn Proteose peptone Bacto-bile salts Sodium thiosulfate Calcium carbonate 5g 1g 30g 10g [ 42 ]

SVTT: Nguyn Thi Trng

Bi Thu Hoch Dung dch Iodine Iodine Potassium Iodine ED

Ngi hng dn: CN. Nguyn Vn Tm

6g 5g 20g

Cn 46g mi trng kh tetrathionate ha tan vo 1 lt nc ct, un si. ngui li 45oC, cho thm 20ml dung dch Iodine, trn u phn phi vo ng nghim v khun, mi ng 10ml. Khng un nng sau khi thm Iodine. Tetrathionate cn bn gi t lnh lu c nhiu ngy. Sau khi cho thm Iodine vo mi trng ch dng trong 1 ngy. 4.1.5. Mi trng phn lp Mi trng thch MC (Mac Conkey Agar) Dng phn lp cc vi khun ng rut ngn chn vi khun Gram dng mc Peptone 17g Proteose peptone 3g Lactose 10g Bile Salt N:3 1,5g NaCl 5g Agar 13,5g Neuture red 0,03g pH = 8 Cn 50g mi trng MC kh ha tan vo 1000ml nc ct. un si cho tan, iu chnh pH = 7,2, kh khun 121oC trong 15 pht. Ly ra ngui khong 50oC phn phi vo hp Petri v khun ch mi trng c li, lt ngc hp thch vo t 37oC. 4.1.6. Cc mi trng sinh ha khc Mi trng Chapman Dng nh danh t cu khun Proteose peptone Beef Extract Mannitol NaCl Agar Phenol Red 10g 1g 10g 75g 20g 0,025g

Cn cc cht theo cng thc. Ha tan vo 1 lt nc ct un si cho tan. iu chnh pH = 7,4 phn phi vo ng nghim 16 x 125ml, mi ng 3ml. hp kh trng 121oC trong 15 pht. Ly ra nghing.

SVTT: Nguyn Thi Trng

[ 43 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Mi trng cn bn Decacboxylase (Decacboxylase medium base) Peptone Yeast Extract Dextrose Brom cresol purple pH = 6,5 5g 3g 1g 0,02g (2ml dung dch 1%)

Cn 9g hn hp trn ha tan vo 1 lt nc ct. un si cho tan, thm L.Lysine, L.Arginine, L.Ornithine theo t l 0,5%. un si iu chnh pH = 6,5 phn phi vo ng nghim 19 x 150mm, mi ng 5ml mi trng. Hp 121oC trong 15 pht. 4.2. Yu cu pha ch Cn ng chnh xc tng thnh phn. Trnh ly nhim t mi trng bn ngoi. Cc mi trng sau khi pha ch, cn phi ct gi trong t lnh.

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[ 44 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Phn 3:

KT QU V THO LUN

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[ 45 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

1. Hnh dng khun lc trn mi trng BA v MC

MC Hnh dng

BA Trn, li

Khng thy xut hin khun lc

Mu sc

Trng c ng theo theo tng chm, hnh dng ging chm nho

Hnh thc

Bin lun Mc ch chnh ca vic cy mu trn mi trng MC v BA: Xem hnh dng khun lc. Xc nh cu khun hay trc khun.

Hnh 3.1: Hnh dng khun lc trn mi trng MC v BA 2. Nhum GRAM Sau khi tin hnh cc bc nhum Gram mu theo qui nh, ta tin hnh soi mu di knh hin vi in t 100x. Kt qu: Vi khun c hnh dng trn, li, bt mu thuc nhum.

SVTT: Nguyn Thi Trng

[ 46 ]

Bi Thu Hoch Cu khun Gram (+)

Ngi hng dn: CN. Nguyn Vn Tm

Hnh 3.2: Hnh nh khun lc di knh hin vi quang trng 100x 3. nh danh Staphylococcus aureus

Th nghim Catalse Oxidase SAULATEX

Kt qu

(+) (-) (+)

Hnh 3.3: Th nghim Catalse (+) v Oxidase (-)

SVTT: Nguyn Thi Trng

[ 47 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Hnh 3.4: Dung dch thuc th SAUlatex v th nghim SAUlatex (+) 4. Kt qu khng sinh mu mu

Hnh 3.5: Kt qu khng sinh trn a

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[ 48 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm Bng 6: Tng kt s liu khng sinh trn 5 a

a Khng Sinh

M Hm Lng (Code)

ng knh vng vi khun o bng mm trn Khng (R) Trung gian (I) Nhy (S) 30 1,5 17 30

Amikacin Ampicillin Amoxlcallin Amoxlcallin/clavuanic acid CEPHALOSPORIN v CEPHEM khc Cephalexin Cefepime Cefoperazone Cefuroxime Cefutriaxone Ceftazidime Cefuroxime acetil

Ak Am Ax Ac

30g 10g 10g 20/10g

Cp Cm Cf Ct Cx Cz Cu

g 30g 75g 30g 30g 30g 30g 9

21 26 28 23 23

23

SVTT: Nguyn Thi Trng

[ 49 ]

Bi Thu Hoch Cefaclor Imipenem Gentamicin Kanamycin Netilmicin Tobramycin Tetracycline Doxyciline Ciprofloxacin Norfloxacin Ofloxacin Nalidixic acid Trirnethoprim/sulfamethoxazole Chloramphenicol Penicicllin Clindamycin Cr Im Ge Kn Ni Tb Te Dx Ci Nr Of Ng Bt Cl Pn cL

Ngi hng dn: CN. Nguyn Vn Tm 30g 10g 10g 30g 30g 10g 30g 30g 5g 10g 5g 30g 1,25/23,75g 30g 10 units 2g 0,7 23 2 3.2 26 23 27 27 23 29 25 33 34 31 30 31

SVTT: Nguyn Thi Trng

[ 50 ]

Bi Thu Hoch Erythromycin Piperacillin Pefloxacin Neomicin Colistin Levofloxacin Er Pt Pf Ne Co Lv

Ngi hng dn: CN. Nguyn Vn Tm 15g 100g 5g 30g 10g 5g 0,4 39 1,2 26 30 25

SVTT: Nguyn Thi Trng

[ 51 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

SVTT: Nguyn Thi Trng

[ 52 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Bnh vin a Khoa Khu Vc Hng Ng l mt bnh vin c uy tn v trch nhim cao trong cng tc y t phc v nhu cu m bo sc khe cho ngi dn. Bnh vin c mt c s trang thit b y v hin i phc v tt cho cng tc y t, bn cnh vi mt i ng cc cn b y t c trch nhim v nng lc cao, Bnh Vin a Khoa Khu Vc Hng Ng lun l mt trong nhng bnh vin c hiu qu v t c nhiu thnh cng ln, v lun l ch da vng chc cho ngi dn. Trong qu trnh thc tp v trc tip tham gia lm vic ti phng Vi Sinh l khong thi gian gip em c c nhng bi hc kinh nghim v b ch, v cng gip em cng c li nhng kin thc m thy c truyn t trn ging ng cho em, ng thi cng gip em bit chng p dng vo thc t nh th no. Ngoi ra, thi gian lm vic ti phng Vi sinh cng gip cho em tm hiu c tc phong lm vic, v k lut lm vic, cch giao tip v quan h gia cc c ch cn b trong mi trng lm vic chuyn nghip. Qua thi gian thc tp ti phng Vi sinh Khoa Xt Nghim Chn on Hnh nh, gip em nm c nhng phng php v cch tip nhn, phn tch mu, c bit l: Thu nhn v x l mu t cc bnh nhn. X l mu ly t cc bnh nhn cc Khoa ca bnh vin. Nng cao kh nng thc hnh x l cc vi sinh vt gy bnh. Tm hiu v vn dng c nhiu cc phn ng sinh ha mi. C thm nhng kin thc mi v kha nng khng khng sinh ca vi sinh vt gy bnh.

Sau qu trnh thc tp, em c nhn xt nh sau: Bnh vin a Khoa Khu Vc Hng Ng l Bnh vin c c s vt cht hin i, vi mt bng rng v cch b tr phng ban mt cch hp l, gip thun tin cho cng vic. Bnh vin c mt b my qun l ng u l Ban gim c y nng lc, ni quy cht ch cng vi lng cn b cng nhn vin c trnh chuyn mn cao, y kinh nghim v nhit tnh trong cng vic. Phng vi sinh c y cc dng c v thit b trong cng tc phn tch v x l bnh phm. Cc c ch trong khoa lun lun hng dn v gip nhit tnh. Bnh vin lun lun c nhng nh hng mi y thnh cng, lun lun tm ti v to ra nhng phng php ti u ha trong vic khm v cha bnh cho nhn dn. Bnh vin lun lun c s quan tm chm sc n i sng v sinh hot ca cc cn b cng chc lm vic, ngoi ra cn tham gia nhiu cng tc x hi khc.

SVTT: Nguyn Thi Trng

[ 53 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

Vn an ton lao ng lun c ch trng u tin hang u, nht l cng tc phng chy cha chy, vi cc trang thit b y v thng xuyn c cc lp tp hun phng chy cha chy cho cc cn b Bnh vin. Lun m bo cho sc khe cho cc cn b v nhn vin Bnh vin, lun lun m nhiu hot ng th thao v vn ha cho ton th. Thi gian thc tp c hn, nn vic c nhng thiu st trong bi bo co l iu khng th trnh khi. V th mong cc thy c trong khoa cng nh cc c ch cn b ca phng Xt Nghim Vi sinh gip v ng gp kin cho bi bo co c hon thin hn. Cui cng, em xin chc Ban gim c Bnh vin a Khoa Khu Vc Hng Ng, cc c ch cn b cng chc, v cc thy c lun c c sc khe, hon thnh cng tc v c nhiu thnh tu trong tng lai.

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[ 54 ]

Bi Thu Hoch Ti liu tham kho

Ngi hng dn: CN. Nguyn Vn Tm

[1]. Vi sinh vt Y hc, 2007 B mn Vi Sinh, Trng i Hc Y H Ni Nh xut bn Y Hc. [2]. Vi Sinh Y Hc Thc Hnh Trng i Hc Y Dc TPHCM Khoa iu Dng K Thut Y B Mn Xt Nghim Nm xut bn 2002. [3]. Vi Khun hc Trng i Hc Y Dc TPHCM Khoa Y B mn Vi Sinh Nm xut bn 2002. [4]. Phng Php Phn Tch Vi Sinh Vt Trong Nc, Thc Phm v M Phm Trn Linh Phc NXB Gio Dc 2009. [5]. Cm Nang Cc K Thut Xt Nghim Vi Sinh Lm Sng Dng Cho Cc Phng Th Nghim Bnh Vin Trng i Hc Y Dc TPHCM TS. Bs. Phm Hng Vn Nm xut bn 2002. [6]. Daksgaad A, et al. J Clin Microbio 1999; 37(3): 734 741. [7]. Medical Microbiology & Immunology, Examination & Board Review Warren Levinson Ernest Jawetz Mc Graw Hill International editions Health Professions Series 2002. [8]. PGSTS Nguyn Hng Tin, PGSTS Bi Minh c, PGSTS Nguyn Vn Dp _ Vi sinh vt thc phm k thut kim tra v ch tiu nh gi an ton thc phm. [9]. Ths Phm Minh Nht, Gio trnh phn tch nh gi cht lng thc phm i Hc K Thut Cng ngh TPHCM, 2010.

Internet [9]. http://textbookofbacteriology.net/themicrobialworld/staph.html.


[10]. http//www.pdb.org/pdb/explore.do?structureId=3SEB.

[11]. www.healthinfotranslations.org/pdfDocs/MRSA_VIET.pdf [12]. http://www.lrc-tnu.edu.vn. [13]. www.yduocngaynay.com [14]. www.google.com.vn

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[ 55 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

NHT K THC TP
Tun s: 01 Cng vic thc hin Th Ngy Sng: Np giy gii thiu ti phng Hnh chnhT Chc. KTV Nguyn Thanh V Tham quan phng xt nghim vi sinh. Chiu: B sung h s thc tp. Ti: Cy dch m o ca sn ph. Sng: Thu mu mu t ngi bnh. X l thng tin ngi bnh. Xem vi khun lao. Chiu: Thu nhn s liu ca a im thc tp Ngi hng dn

Hai

12/3

CN. Nguyn Vn Tm.

Ba

13/3

14/3

Sng: Thu thp s liu mu. Kim tra mu. Kim tra vi sinh vt mc trn a (mu ly CN. Nguyn Vn Tm. t cc bnh nhn). un mi trng Quan st qu trnh thc hin khng sinh t mu dch bnh. Chiu: Quan st quy trnh xc nh Staphylococus Aureus trn mu dch bnh. Sng: c kt qu mc khng khun Staphylococus Aureus. o ng knh khng khun. Chiu: V sinh dng c.

Nm

15/3

CN. Nguyn Vn Tm.

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[ 56 ]

Bi Thu Hoch Lc mi trng 2 pha. Kim tra mu.

Ngi hng dn: CN. Nguyn Vn Tm

Su

16/3

Sng: Cy vi sinh vt vo mi trng BA v MC. CN.Nguyn Vn Tm Chiu: Chun b mi trng, un mi trng. KTV.Nguyn Thanh V Cy mu dch sn ph. Ti: Cy dch m h sn ph. Thu nhn mu mu. Sng: Kim tra mu. un mi trng MC v MHA. Chiu: Cy mu dch vo mi trng MC v BA

By

17/3

CN.Nguyn Vn Tm

CN

18/3

Tun s: 02 Cng vic thc hin Th Ngy Sng: Xem kt qu cy mu. Nhn xt kt qu. Lm phin nhum Gram c kt qu trn knh hin vi. Lm khng sinh cho mu. Chiu: Kim tra mu. Ph vic cho phng xt nghim. Sng: Kim tra mu. Th nghim nhum Gram trn mu mu. Chiu: Kim tra kt qu cy mu dch v mu. Tng hp ti liu c lien quan Sng: Xem kt qu mu cy.

Ngi hng dn

Hai

19/3

CN. Nguyn Vn Tm

Ba

20/3

CN. Nguyn Vn Tm

21/3

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[ 57 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm CN. Nguyn Vn Tm

c khng sinh mu dch v mu mu. Chiu: Tng hp ti liu bo co. Cy mu dch t cung ca sn ph. Sng: c kt qu mu cy. Chiu: Tng sinh mu mu. Cy chuyn. Sng: Cy mu. Kim tra mu tng sinh. Lm cc th nghim sinh ha. Chiu: un mi trng MC v MHI Lm khng sinh . Sng: c kt qu khng sinh . Kim tra kt qu cc th nghim sinh ha. Kim tra mu cy mu.

Nm

22/3

CN. Nguyn Vn Tm

Su

23/3

CN. Nguyn Vn Tm

By

24/3

CN. Nguyn Vn Tm

CN

25/3

Tun s: 02 Cng vic thc hin Th Ngy Sng: c kt qu khng sinh . Xem xt kt qu cc th nghim sinh ha nhm trc khun Gram (-). Chiu: Cng vic trng. Sng: Thu nhn kt qu. Chiu: Tng kt s liu.

Ngi hng dn

Hai

26/3

CN. Nguyn Vn Tm

Ba

27/3

CN. Nguyn Vn Tm

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[ 58 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm Thc hin bo co thc tp

28/3 CN. Nguyn Vn Tm In bo co thc tp. Np bo co thc tp cho phng T Chc Hnh Chnh, Trng Khoa v Cn b hng dn

Nm

29/3

CN. Nguyn Vn Tm

Su By CN

30/3 31/3 01/4

SVTT: Nguyn Thi Trng

[ 59 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm MC LC

LI CM N DANH MC HNH DANH MC BNG S LI M U PHN 1: TNG QUAN CHNG I: I NT V BNH VIN A KHOA KHU VC HNG NG 1.1. Lch S Hnh Thnh Bnh Vin ......................................................................... 4 1.2. Ban Lnh o .................................................................................................... 6 1.3. Khoa Xt Nghim .............................................................................................. 8 1.4. Cn B Cng Nhn Vin Khoa Chn on Hnh nh Xt Nghim ................ 8 1.5. Chc nng Nhim V ..................................................................................... 8 1.6. Cc Thit B Khoa Chn on Hnh nh Xt Nghim.................................... 8 CHNG II: TNG QUAN V VI KHUN STAPHYLOCOCCUS AUREUS 2.1. T cu vng Staphylococcus aureus ................................................................... 13 2.1.1. Lch s pht hin............................................................................................. 13 2.1.2. c im phn loi ......................................................................................... 13 2.1.3. c im sinh ha........................................................................................... 14 2.1.4. c im vi khun hc.................................................................................... 14 2.1.5. H gen t cu vng Staphylococcus aureus ..................................................... 16 2.2. Ni c t rut staphylococcal enterotoxin B ..................................................... 16 2.2.1. Cu trc phn t staphylococcal enterotoxin B................................................ 16 2.2.2. C ch gy c ca staphylococcal enterotoxin B ........................................... 17 2.3. Thc trng nhim Staphylococcus aureus Vit Nam ....................................... 17 2.4. Phng nga, iu tr v x l bnh...................................................................... 17 2.4.1. Phng nga ..................................................................................................... 17 2.4.2. iu tr............................................................................................................ 18 2.4.3. X l bnh ....................................................................................................... 18 PHN 2: VT LIU V PHNG PHP

SVTT: Nguyn Thi Trng

[ 60 ]

Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

1.1. Phng php ly v gi bnh phm ................................................................... 21 1.1.1. Thi gian v a im th nghim .................................................................... 21 1.1.2. i tng th nghim ...................................................................................... 21 1.2. Cch ly bnh phm ........................................................................................... 21 1.3. Cch gi bnh phm .......................................................................................... 22 2.1. Cy mu ............................................................................................................ 25 2.1.1. Ch nh cy mu ............................................................................................ 25 2.1.2. Thi im cy mu ......................................................................................... 25 2.1.3. Cch ly mu cy ....................................................................................... 25 2.1.4. Mi trng cy mu ........................................................................................ 25 2.2. Theo di cy mu............................................................................................... 25 2.3. Vn vi khun ngoi nhim ............................................................................. 26 3.1. Cc th nghim sinh ha nh danh ................................................................... 30 3.1.1. Th nhim -Lactamase .................................................................................. 30 3.1.2. Trc nghim Catalase ...................................................................................... 31 3.1.3. Kh nng tng trng v ln men .................................................................... 32 trn mi trng M.S.A (Chapman) 3.1.4. Trc nghim Coagulase ................................................................................... 32 3.1.5. Trc nghim Novobiocin................................................................................. 32 3.1.6. Th nghim SAUTlATEX (STAPHYLATEX) ............................................... 33 3.2. Phng php nhum Gram ................................................................................ 34 3.2.1. Nguyn tc ...................................................................................................... 34 3.2.2. Dng c v thuc nhum ................................................................................ 34 3.2.3. K thut .......................................................................................................... 34 3.2.4. Kt qu ........................................................................................................... 35 3.3. K thut lm khng sinh ................................................................................ 35 3.3.1. Nguyn tc ...................................................................................................... 36 3.3.2. Chun b ......................................................................................................... 36 3.3.3. K thut .......................................................................................................... 36 4.1. Cch pha ch mt s mi trng nui cy vi khun ........................................... 40

SVTT: Nguyn Thi Trng

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Bi Thu Hoch

Ngi hng dn: CN. Nguyn Vn Tm

4.1.1. Mi trng dinh dng c bn........................................................................ 40 4.1.2. Mi trng b ................................................................................................. 41 4.1.3. Mi trng chuyn ch ................................................................................... 42 4.1.4. Mi trng phong ph ha .............................................................................. 42 4.1.5. Mi trng phn lp ....................................................................................... 43 4.1.6. Cc mi trng sinh ha khc ......................................................................... 43 4.2. Yu cu pha ch................................................................................................. 44 PHN 3: KT QU V THO LUN 1. Hnh dng khun lc trn mi trng BA v MC .................................................. 46 2. Nhum Gram ........................................................................................................ 46 3. nh danh Staphylococcus aureus ........................................................................ 47 4. Kt qu khng sinh mu mu ........................................................................... 48 Ti liu tham kho .................................................................................................. 55 Internet.................................................................................................................... 55 NHT K THC TP ........................................................................................... 56 MC LC ............................................................................................................... 60

SVTT: Nguyn Thi Trng

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