1株泛耐药肺炎克雷伯菌质粒介导耐药的机制研究
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Investigation on the mechanism of plasmid-mediated resistance in one strain of pandrug resistant Klebsiella pneumonia
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    摘要:

    摘要:目的:观察质粒与1株泛耐药肺炎克雷伯菌(PRKP)耐药机制间的关系。 方法:用纸片扩散法和微量肉汤稀释法对1株临床分离的PRKP进行药敏试验。用PCR检测PRKP携带的碳青霉烯酶基因GES-1~9、GES-11、OXA-48-like、IMP、VIM、KPC-1~5和NDM,以及广宿主质粒IncN家族的复制酶基因repA。提取该菌株质粒并行琼脂糖凝胶电泳,进行质粒接合、转化试验,PCR检测转化接合子碳青霉烯酶基因和repA。 结果:从PRKP菌株分离到pIncN-XM和pKPC-XM质粒,前者可经接合转移到E.coli J53,PCR检测显示其为一种具广宿主质粒IncN特征的质粒,并携带喹诺酮类抗性基因;后者接合试验阴性,但可经转化导入感受态细菌E.coli DH5α,携带blaKPC-2型基因。 结论: 该PRKP菌株泛耐药的机制与其含携带blaKPC-2型基因的质粒和携带喹诺酮类抗性基因并具有广宿主质粒IncN特征的质粒密切相关。

    Abstract:

    Abstract: Objective: To investigate the mechanism of plasmid-mediated resistance in one strain of pandrug resistant Klebsiella pneumonia (PRKP). Methods: The susceptibility of the PRKP strain was determined by the disk diffusion method and the automated microbroth dilution. PCR was used to identify the carbapenemase genes, including GES-1~GES-9, GES-11, OXA-48-like, IMP, VIM, KPC-1~KPC-5 and NDM, and the replicase gene repA of broad-host-range plasmid IncN in the PRKP strain. Then, the plasmid in the PRKP strain was extracted, and isolated by agarose gel electrophoresis. After the conjugation and transformation experiments were performed, the carbapenemase genes and repA in the conjugant were detected by PCR. Results: The pIncN-XM and pKPC-XM plasmids were isolated from the PRKP strain. The pIncN-XM could be transferred into E. coli J53 by the conjugation, which was an IncN-like plasmid and carried quinolone-resistant genes. However, the pKPC-XM carrying the bla>sub>KPC-2 gene was unable to be transferred into E. coli J53 but into E. coli DH5α by the transformation. Conclusion: The pandrug resistance mechanism of the PRKP may be closely related to the presence of two harbored plasmids, one carrying bla>sub>KPC-2 gene, and the other carrying quinolone-resistant genes and with IncN like feature.

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张国强,姚艺辉,余晓露.1株泛耐药肺炎克雷伯菌质粒介导耐药的机制研究[J].临床检验杂志,2013,31(10):788-791

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  • 收稿日期:2013-07-30
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  • 在线发布日期: 2013-12-11
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