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霍乱弧菌多位点可变数目串联重复序列分析与脉冲场凝胶电泳及毒力分型的比较

Multiple-locus variable-number tandem repeat analysis of Vibrio cholerae in comparison with pulsed field gel electrophoresis and virulotyping.

作者信息

Teh Cindy Shuan Ju, Chua Kek Heng, Thong Kwai Lin

机构信息

Faculty of Science, Institute of Biological Sciences, University of Malaya, 50603 Kuala Lumpur, Malaysia.

出版信息

J Biomed Biotechnol. 2010;2010:817190. doi: 10.1155/2010/817190. Epub 2010 Jun 30.

Abstract

Molecular analysis of Malaysian Vibrio cholerae was carried out using a multiple-locus variable-number tandem repeat analysis (MLVA) assay based on 7 loci of V. cholerae. The discriminatory ability of the assay was compared with pulsed-field gel electrophoresis (PFGE) using 43 Malaysian V. cholerae isolated from various sources. In addition, the virulotypes of the strains were determined. Based on MLVA, 38 allelic profiles were obtained (F = 0.63) while PFGE generated 35 pulsotypes (F = 0.71). Simpson's index of diversity for different VNTR loci ranged from 0.59 to 0.92. The combined loci increased the discriminatory index to 0.99 which was comparable with PFGE (D = 0.99). Most of the environmental non-O1/non-O139 strains harbored rtxA, rstR, toxR, and hlyA only, and the virulotype of this serogroup was significantly different (P < .01) from clinical/environmental O1 and environmental O139 strains. In conclusion, the MLVA assay developed in this study was a useful genotyping tool with comparable discriminatory power with PFGE. In addition, the combination of the two approaches can further distinguish the strains from different sources and geographical regions of isolation.

摘要

采用基于霍乱弧菌7个位点的多位点可变数目串联重复序列分析(MLVA)方法,对马来西亚霍乱弧菌进行分子分析。利用从不同来源分离得到的43株马来西亚霍乱弧菌,将该分析方法的鉴别能力与脉冲场凝胶电泳(PFGE)进行比较。此外,还确定了菌株的毒力型。基于MLVA,获得了38个等位基因谱(F = 0.63),而PFGE产生了35个脉冲型(F = 0.71)。不同VNTR位点的辛普森多样性指数在0.59至0.92之间。组合位点将鉴别指数提高到0.99,与PFGE相当(D = 0.99)。大多数环境非O1/非O139菌株仅携带rtxA、rstR、toxR和hlyA,该血清群的毒力型与临床/环境O1和环境O139菌株显著不同(P <.01)。总之,本研究开发的MLVA分析方法是一种有用的基因分型工具,其鉴别能力与PFGE相当。此外,两种方法的结合可以进一步区分来自不同分离来源和地理区域的菌株。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d156/2910556/5d33e8d7c3f2/JBB2010-817190.001.jpg

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