一种用于区分空肠弯曲菌和结肠弯曲菌的聚合酶链反应与限制性内切酶联合检测法。

A combined polymerase chain reaction and restriction endonuclease enzyme assay for discriminating between Campylobacter coli and Campylobacter jejuni.

作者信息

Comi G, Pipan C, Botta G, Cocolin L, Cantoni C, Manzano M

机构信息

Dipartimento di Scienze degli Alimenti, Facoltà di Agraria, Università di Udine, Italy.

出版信息

FEMS Immunol Med Microbiol. 1996 Nov;16(1):45-9. doi: 10.1111/j.1574-695X.1996.tb00110.x.

DOI:10.1111/j.1574-695X.1996.tb00110.x

PMID:8954352

Abstract

A combined polymerase chain reaction and restriction endonuclease (RE) enzyme assay was developed to discriminate between Campylobacter coli and Campylobacter jejuni. Amplimers of the FlaA gene obtained by PCR were digested with AluI and HinfI to distinguish C. coli from C. jejuni. With AluI digestion C. jejuni-specific bands were observed at 110, 140 and 160 bp and C. coli-specific bands at 293 and 147 bp. C. jejuni-specific bands of 349 and 109 bp were found by HinfI digestion but HinfI did not digest the FlaA amplimer of C. coli. This combined technique is fast and easy to perform, and distinguishes the two campylobacters unequivocally.

摘要

开发了一种聚合酶链反应和限制性内切酶（RE）酶联分析法，以区分空肠弯曲菌和结肠弯曲菌。通过PCR获得的FlaA基因扩增子用AluI和HinfI进行消化，以区分结肠弯曲菌和空肠弯曲菌。用AluI消化后，在110、140和160 bp处观察到空肠弯曲菌特异性条带，在293和147 bp处观察到结肠弯曲菌特异性条带。通过HinfI消化发现了349和109 bp的空肠弯曲菌特异性条带，但HinfI未消化结肠弯曲菌的FlaA扩增子。这种联合技术快速且易于操作，能明确区分这两种弯曲菌。