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利用聚合酶链反应对表面蛋白基因重复区进行快速分型新生儿表皮葡萄球菌分离株。

Rapid typing of neonatal Staphylococcus epidermidis isolates using polymerase chain reaction for repeat regions in surface protein genes.

机构信息

Department of Paediatrics, Orebro University Hospital, 701 85, Orebro, Sweden.

出版信息

Eur J Clin Microbiol Infect Dis. 2010 Jun;29(6):699-704. doi: 10.1007/s10096-010-0917-z. Epub 2010 Apr 11.

Abstract

Staphylococcus epidermidis is a significant pathogen in neonatal sepsis and other nosocomial infections. For further investigations of the colonisation patterns and invasive pathways, typing methods that are applicable on large populations of bacterial isolates are warranted. In the present study, a genotyping method based on polymerase chain reaction (PCR) for the repeat regions of four genes (sdrG, sdrF, aap and sesE) that encode for bacterial surface proteins was developed and applied to a sample of well-characterised neonatal blood isolates of S. epidermidis (n = 49). The PCR products were visualised on agarose gel (sdrG, sdrF and sesE) or by fragment analysis (aap). The discriminatory index (D-index) for genotyping of the different genes was compared to genotyping by pulsed-field gel electrophoresis (PFGE). The highest D-index for the PCR-based typing methods was found for the combination of sdrF, sdrG and aap (D-index 0.94), whereas the optimal two-gene combination (sdrF and aap) resulted in a D-index of 0.92. We conclude that the described method can be used for the genotyping of large populations of S. epidermidis isolates with a sufficient discriminatory capacity, and we suggest that the combination of sdrF and aap is the most suitable to use.

摘要

表皮葡萄球菌是新生儿败血症和其他医院感染的重要病原体。为了进一步研究定植模式和侵袭途径,需要应用于大量细菌分离株的分型方法。在本研究中,开发了一种基于聚合酶链反应(PCR)的基因分型方法,用于编码细菌表面蛋白的四个基因(sdrG、sdrF、aap 和 sesE)的重复区域,并将其应用于一组特征明确的表皮葡萄球菌新生儿血分离株(n=49)。PCR 产物在琼脂糖凝胶上可视化(sdrG、sdrF 和 sesE)或通过片段分析(aap)进行可视化。不同基因的基因分型的鉴别指数(D-index)与脉冲场凝胶电泳(PFGE)的基因分型进行了比较。基于 PCR 的分型方法中,sdrF、sdrG 和 aap 的组合具有最高的 D-index(0.94),而最佳的双基因组合(sdrF 和 aap)则产生 0.92 的 D-index。我们得出结论,该方法可用于具有足够鉴别能力的大量表皮葡萄球菌分离株的基因分型,并且我们建议 sdrF 和 aap 的组合是最适合使用的。

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