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四种方法(包括半自动化重复扩增多态性 PCR)用于肠球菌粪肠球菌耐万古霉素株的分型比较。

Comparison of four methods, including semi-automated rep-PCR, for the typing of vancomycin-resistant Enterococcus faecium.

机构信息

Centre National de Référence de la Résistance aux Antibiotiques, Service de Microbiologie, CHU Côte de Nacre, Université de Caen, avenue Côte de Nacre, 14033 Caen Cedex, France.

出版信息

J Microbiol Methods. 2011 Jan;84(1):74-80. doi: 10.1016/j.mimet.2010.10.014. Epub 2010 Nov 6.

Abstract

We have assessed the performance of semi-automated rep-PCR (Diversilab®) and multilocus sequence typing (MLST) in comparison to pulsed-field gel electrophoresis (PFGE) for typing a collection of 29 epidemiologically characterized vancomycin-resistant Enterococcus faecium (VRE). Sixteen strains that harbored the Tn1546 element were typed by PCR mapping. The discriminative power of the typing methods was calculated by the Simpson's index of diversity, and the concordance between methods was evaluated by the Kendall's coefficient of concordance. Semi-automated rep-PCR appeared as discriminative as PFGE and was further compared with PFGE for typing 67 VRE isolated during a hospital outbreak. Rep-PCR appeared to be more discriminative than PFGE for this second set of strains. Reproducibility of DiversiLab® was also tested against 35 selected isolates. Only three showed less than 97% similarity, indicating high reproducibility at this level of discrimination. In conclusion, semi-automated rep-PCR is a useful tool for rapid screening of VRE isolates during an outbreak, although cost of the system may be limiting for routine implementation. PFGE, which remains the reference method, should be used for confirmation and evaluation of the genetic relatedness of epidemic isolates.

摘要

我们评估了半自动化重复扩增多态性 DNA 分析(Diversilab®)和多位点序列分型(MLST)与脉冲场凝胶电泳(PFGE)在 29 株经流行病学特征鉴定的万古霉素耐药粪肠球菌(VRE)分型方面的性能。16 株携带 Tn1546 元件的菌株通过 PCR 图谱进行了分型。通过 Simpson 多样性指数计算了分型方法的辨别力,并通过 Kendall 协和系数评估了方法之间的一致性。半自动化重复扩增多态性 DNA 分析与 PFGE 一样具有辨别力,并进一步用于对医院爆发期间分离的 67 株 VRE 进行分型。对于这第二组菌株,Rep-PCR 似乎比 PFGE 更具辨别力。还针对 35 株选定的分离株测试了 DiversiLab®的重现性。只有 3 株的相似度低于 97%,表明在这种分辨力水平下具有很高的重现性。总之,半自动化重复扩增多态性 DNA 分析是在爆发期间快速筛选 VRE 分离株的有用工具,尽管系统的成本可能限制了其常规应用。PFGE 仍然是参考方法,应用于确认和评估流行株的遗传相关性。

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