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开发一种基于细胞致死扩张毒素(cdt)基因的种特异性多重PCR检测方法,用于检测和鉴定空肠弯曲菌、结肠弯曲菌和胎儿弯曲菌。

Development of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the detection and identification of Campylobacter jejuni, Campylobacter coli and Campylobacter fetus.

作者信息

Asakura Masahiro, Samosornsuk Worada, Hinenoya Atsushi, Misawa Naoaki, Nishimura Kazuhiko, Matsuhisa Akio, Yamasaki Shinji

机构信息

Graduate School of Life and Environmental Sciences, Osaka Prefecture University, Osaka, Japan.

出版信息

FEMS Immunol Med Microbiol. 2008 Mar;52(2):260-6. doi: 10.1111/j.1574-695X.2007.00369.x. Epub 2008 Jan 31.

Abstract

A cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the detection of cdtA, cdtB or cdtC gene of Campylobacter jejuni, Campylobacter coli or Campylobacter fetus, respectively, was developed and evaluated with 76 Campylobacter strains belonging to seven different species and 131 other bacterial strains of eight different genera. The cdtA, cdtB or cdtC gene of C. jejuni, C. coli or C. fetus, respectively, could be successfully amplified using the corresponding set of primers in a highly species-specific manner. Furthermore, the specific primer set for the cdtA, cdtB or cdtC gene of a particular species could amplify the desired gene from a mixture of DNA templates of any of two or all three species. The detection limit of C. jejuni, C. coli or C. fetus was 10-100 CFU tube(-1) by the multiplex PCR assay on the basis of the presence of the cdtA, cdtB or cdtC gene. These data indicate that the cdt gene-based multiplex PCR assay may be useful for rapid and accurate detection as well as identification of Campylobacter strains in a species-specific manner.

摘要

开发了一种基于细胞致死膨胀毒素(cdt)基因的种特异性多重PCR检测方法,用于分别检测空肠弯曲菌、结肠弯曲菌或胎儿弯曲菌的cdtA、cdtB或cdtC基因,并对76株属于7个不同种的弯曲菌菌株和131株属于8个不同属的其他细菌菌株进行了评估。使用相应的引物组,可分别以高度种特异性的方式成功扩增空肠弯曲菌、结肠弯曲菌或胎儿弯曲菌的cdtA、cdtB或cdtC基因。此外,针对特定种的cdtA、cdtB或cdtC基因的特异性引物组可从任意两种或所有三种菌的DNA模板混合物中扩增出所需基因。基于cdtA、cdtB或cdtC基因的存在,多重PCR检测法对空肠弯曲菌、结肠弯曲菌或胎儿弯曲菌的检测限为10 - 100 CFU/管。这些数据表明,基于cdt基因的多重PCR检测法可能有助于以种特异性方式快速、准确地检测和鉴定弯曲菌菌株。

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