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首页> 外文期刊>Journal of Clinical Microbiology >Molecular Typing of Methicillin-Resistant Staphylococcus aureus: Can PCR Replace Pulsed-Field Gel Electrophoresis?
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Molecular Typing of Methicillin-Resistant Staphylococcus aureus: Can PCR Replace Pulsed-Field Gel Electrophoresis?

机译:耐甲氧西林金黄色葡萄球菌的分子分型:PCR可以代替脉冲场凝胶电泳吗?

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摘要

Pulsed-field gel electrophoresis (PFGE) is considered the “gold standard” for molecular typing of methicillin-resistant Staphylococcus aureus (MRSA). However, the method is time-consuming and expensive, and its discriminatory power may not be necessary in outbreak situations. We used a rapid multiplex PCR-based method with published primers and compared the results with those obtained by PFGE. A total of 75 clinical isolates were typed: 59 strains originated from our prospectively collected clinical strains and were epidemiologically unrelated; 16 strains came from an outbreak that was epidemiologically well defined in time and space. A primer mix of the spa gene, the coa gene, and the hypervariable region adjacent to mecA gene was used for multiplex PCR. Both PFGE and PCR clustered the 75 strains into 41 different genotypes. Concordance of the results was 100% for strains originating from the outbreak. Overall, both methods produced concordant results in 72% of cases. A total of 16% were clustered together by PFGE, but not by PCR and 12% were clustered together by PCR but not by PFGE, respectively. The turnaround time was only 8 h for PCR but 5 days for PFGE. This PCR-based method is excellent for rapid and inexpensive typing of MRSA in an outbreak setting, but the discriminatory power and reproducibility are still insufficient to replace PFGE in longitudinal studies in the endemic setting.
机译:脉冲场凝胶电泳(PFGE)被认为是耐甲氧西林金黄色葡萄球菌(MRSA)分子分型的“金标准”。但是,该方法既耗时又昂贵,并且在爆发情况下可能不需要区分力。我们使用了基于快速多重PCR的方法和已发表的引物,并将结果与​​PFGE获得的结果进行了比较。共分出75种临床分离株:59株源自我们前瞻性收集的临床株,在流行病学上无关。在流行病学上在时间和空间上已明确定义的一次疫情暴发了16株。使用 spa 基因, coa 基因和与 mec A基因相邻的高变区的引物混合物进行多重PCR。 PFGE和PCR都将75个菌株聚类为41种不同的基因型。对于源自暴发的菌株,结果的一致性为100%。总体而言,这两种方法在72%的病例中产生了一致的结果。 PFGE分别将16%的聚类在一起,但PCR而不是PFGE将12%的聚类在一起。 PCR的周转时间仅为8 h,而PFGE的周转时间为5天。这种基于PCR的方法非常适合在爆发环境中快速,廉价地分型MRSA,但在地方性环境的纵向研究中,鉴别力和可重复性仍不足以取代PFGE。

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