大肠埃希菌连续分离株氨基糖苷类修饰酶aac_6_b基因分型研究

中图分类号:R378.2+1 文献标识码:A

大肠埃希菌连续分离株氨基糖苷类修饰酶aac(6 ) b 基因分型研究

王 伟1,糜祖煌2,毛剑峰1,徐伟珍1(1.丽水市人民医院,浙江丽水323000;2.无锡市克隆遗传技术研究所)

摘要:目的 了解临床连续分离的大肠埃希菌(ECO)氨基糖苷类饰酶aac(6 ) Ib基因及亚型存在状况。方法 测定临床分离的60株ECO菌对19种抗菌药物的敏感性,采用PCR技术检测氨基糖苷类修饰酶aac(6 ) Ib基因。结果 60株ECO菌连续分离株中aac(6 ) Ib基因阳性11株(18.3%),经DNA测序,2株为aac(6 ) Ib经典型,6株为aac(6 ) Ib C r型,另有2株为新亚型。结论 检出氨基糖苷修饰酶aac(6 ) Ib基因新亚型,在一家医院ECO菌中同时检出aac(6 ) Ib型和aac(6 ) Ib C r 型及新亚型三种亚型为国内首次报道。

关键词:大肠埃希菌;氨基糖苷类修饰酶;aac(6 ) Ib基因;亚型

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D etection and i dentification of clinical pathogenic bacteri a w ith16S and

23S rDNA based oligonucleoti de m icroarray

XING Jian m i n g1,Z HANG Su1,Z HANG H ong he2,SHEN Cu i fen3,B I Dan1,L IG ang1,YAO L i hu i1,WU X i ao fang4(1.D epart ment o f L aboratory M ed ici ne,H uz houM a tern ity and Chil d Care H osp ita,l H uz hou313000,Ch i na)

Abstrac t:O bjective To deve lop a16S rDNA and23S r DNA based o li gonucleoti de m i c roarray f o r detecti on and i dentifica ti on of c li n ical pathogenic bacte ria.M ethod s D ouble PCR was appli ed to amp lify t he segm ents of16S r DNA and23S rDNA genes of t he ta rget bacter i a.A n o li gonucleoti de m icroarray w as constructed to si m u ltaneousl y detect EHEC O157!H7,V ibrio parahae mo l y ticus, Sai monella sp.,V ibr io cholerae,L ister i a m onocy togenes,Ca mpy lobacter jejuni and Shigella sp.T he specific it y,sensitiv it y and repro duc i b ilit y o f the m icroa rray w ere eva l uated.A nd the m icroarray w as used to de tect t he m icrobes in stool speci m ens fro m81pa ti entsw ith d i arrhea and vom iting.Re s u lts The doub l e PCR cou l d si m u ltaneously a m plif y t he target sequences o f16S r DNA and23S rDNA genes o f the seven pat hogens.The sensiti v ity o f the deve l oped o li gonuc l eo tide m icro array w as103cf u/m l and no positive resu lts we re obse rved for non target bacter i a.A nd the coe fficients of var i a ti on in one lot o r among d ifferent l o ts of t he m icro array sli des we re3.89%~ 5 81%.T he positi v e detection ra te o f t he stool spec i m ens by o li gonuc l eotide m i croarray was39.5%(32/81),w ith a co i nc i dence of 96.3%(78/81)for t he pa ti ents and ano t her co i ncidence o f96.8%(31/32)for bac terial g enus or spec i es i dentificati on co m pared to the resu lts by the routi ne bacter i o l og ical exam i na ti ons.Conc l u sion T he16S and23S r DNA based o li gonucleoti de m icroarray to detect the seven pat hogenic bacte ria hasm any advantag es ove r rou tine bac teriolog i ca l exa m i nation such as conven i ent,rapid,accurate,stab le and h i gh throughout w hich is su itab le for c li n ica l speci m en exam i na ti on and ep i dem i o l og ica l field i nvesti ga ti on.

K ey word s:O ligonucleo ti de m icroarray;Pa t hogen i c bacter i a;D etecti on

(收稿日期:2008 03 26,修回日期:2008 06 12)

(本文编辑:王伯昌)