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评估四种分子分型方法作为确定分类学关系和鉴定耶尔森氏菌分离株中物种的工具。

Evaluation of four molecular typing methodologies as tools for determining taxonomy relations and for identifying species among Yersinia isolates.

机构信息

Departamento de Análises Clínicas, Faculdade de Ciências Farmacêuticas de Ribeirão Preto, USP, Ribeirão Preto, SP, Brazil.

出版信息

J Microbiol Methods. 2010 Aug;82(2):141-50. doi: 10.1016/j.mimet.2010.05.005. Epub 2010 May 19.

Abstract

In the last few decades, molecular typing has become an important tool in taxonomic, phylogenetic and identification studies of numerous groups of bacteria, including the yersiniae. In this study, Enterobacterial Repetitive Intergenic Consensus PCR (ERIC-PCR), Pulsed-Field Gel Electrophoresis (PFGE), 16S rRNA gene sequencing and Multilocus Sequence Analysis (MLSA) were performed to determine the ability of these techniques to be used in taxonomy and identification of Yersinia strains. A total of 60 Yersinia strains were genotyped by ERIC-PCR and PFGE. Moreover, an in silico analysis was carried out for 16S rRNA gene sequencing and MLSA, using 68 and 49 Yersinia strains, respectively. A phylogenetic tree constructed from the ERIC-PCR, 16S rRNA gene sequencing and MLSA data grouped most of the Yersinia species into distinct species-specific clusters. In the PFGE assay these clusters were not observed. On this basis, ERIC-PCR, 16S rRNA gene sequencing and MLSA seem to be valuable techniques for use in taxonomic and identification studies of the genus Yersinia, whereas PFGE does not. Furthermore, ERIC-PCR has the advantage of being a cheaper, easier and faster assay than 16S rRNA gene sequencing or MLSA, and for these reasons can be considerate an alternative tool in taxonomic studies of yersiniae.

摘要

在过去的几十年中,分子分型已成为分类学、系统发育学和许多细菌群体鉴定研究的重要工具,包括耶尔森氏菌。在这项研究中,肠杆菌重复基因间一致性聚合酶链反应(ERIC-PCR)、脉冲场凝胶电泳(PFGE)、16S rRNA 基因测序和多位点序列分析(MLSA)被用于确定这些技术在耶尔森氏菌菌株的分类学和鉴定中的应用能力。总共对 60 株耶尔森氏菌进行了 ERIC-PCR 和 PFGE 基因分型。此外,分别对 68 株和 49 株耶尔森氏菌进行了 16S rRNA 基因测序和 MLSA 的计算机分析。基于 ERIC-PCR、16S rRNA 基因测序和 MLSA 数据构建的系统发育树将大多数耶尔森氏菌物种分为不同的种特异性聚类。在 PFGE 检测中没有观察到这些聚类。在此基础上,ERIC-PCR、16S rRNA 基因测序和 MLSA 似乎是用于耶尔森氏菌属分类学和鉴定研究的有价值的技术,而 PFGE 则不然。此外,ERIC-PCR 具有比 16S rRNA 基因测序或 MLSA 更便宜、更容易、更快的优势,因此可以考虑将其作为耶尔森氏菌分类学研究的替代工具。

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