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采用基于聚合酶链式反应(PCR)的方法对从菲律宾分离出的副溶血性弧菌菌株进行分子分型。

Molecular typing of Vibrio parahaemolyticus strains isolated from the Philippines by PCR-based methods.

作者信息

Maluping R P, Ravelo C, Lavilla-Pitogo C R, Krovacek K, Romalde J L

机构信息

Department of Biomedical Sciences and Veterinary Public Health, Faculty of Veterinary Medicine and Animal Science, SLU, Uppsala, Sweden.

出版信息

J Appl Microbiol. 2005;99(2):383-91. doi: 10.1111/j.1365-2672.2005.02571.x.

Abstract

AIM

The main aim of the present study was to use three PCR-based techniques for the analysis of genetic variability among Vibrio parahaemolyticus strains isolated from the Philippines.

METHODS AND RESULTS

Seventeen strains of V. parahaemolyticus isolated from shrimps (Penaeus monodon) and from the environments where these shrimps are being cultivated were analysed by random amplified polymorphic DNA PCR (RAPD-PCR), enterobacterial repetitive intergenic consensus sequence PCR (ERIC-PCR) and repetitive extragenic palindromic PCR (REP-PCR). The results of this work have demonstrated genetic variability within the V. parahaemolyticus strains that were isolated from the Philippines. In addition, RAPD, ERIC and REP-PCR are suitable rapid typing methods for V. parahaemolyticus. All three methods have good discriminative ability and can be used as a rapid means of comparing V. parahaemolyticus strains for epidemiological investigation. Based on the results of this study, we could say that REP-PCR is inferior to RAPD and ERIC-PCR owing to the fact that it is less reproducible. Moreover, the REP-PCR analysis yielded a relatively small number of products. This may suggests that the REP sequences may not be widely distributed in the V. parahaemolyticus genome.

CONCLUSIONS

Genetic variability within V. parahaemolyticus strains isolated in the Philippines has been demonstrated. The presence of ERIC and REP sequences in the genome of this bacterial species was confirmed.

SIGNIFICANCE AND IMPACT OF THE STUDY

The RAPD, ERIC and REP-PCR techniques are useful methods for molecular typing of V. parahaemolyticus strains. To our knowledge this is the first study of this kind carried out on V. parahaemolyticus strains isolated from the Philippines.

摘要

目的

本研究的主要目的是使用三种基于聚合酶链反应(PCR)的技术来分析从菲律宾分离出的副溶血性弧菌菌株之间的遗传变异性。

方法与结果

通过随机扩增多态性DNA聚合酶链反应(RAPD-PCR)、肠杆菌重复基因间共有序列聚合酶链反应(ERIC-PCR)和重复外显子回文聚合酶链反应(REP-PCR),对从虾(斑节对虾)及其养殖环境中分离出的17株副溶血性弧菌进行了分析。这项工作的结果表明,从菲律宾分离出的副溶血性弧菌菌株存在遗传变异性。此外,RAPD、ERIC和REP-PCR是适用于副溶血性弧菌的快速分型方法。这三种方法都具有良好的鉴别能力,可作为比较副溶血性弧菌菌株进行流行病学调查的快速手段。基于本研究结果,我们可以说REP-PCR不如RAPD和ERIC-PCR,因为它的可重复性较差。此外,REP-PCR分析产生的产物数量相对较少。这可能表明REP序列在副溶血性弧菌基因组中分布不广泛。

结论

已证明在菲律宾分离出的副溶血性弧菌菌株存在遗传变异性。证实了该细菌物种基因组中存在ERIC和REP序列。

研究的意义和影响

RAPD、ERIC和REP-PCR技术是副溶血性弧菌菌株分子分型的有用方法。据我们所知,这是首次对从菲律宾分离出的副溶血性弧菌菌株进行此类研究。

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