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通过限制性片段长度多态性分析对空肠弯曲菌鞭毛蛋白基因进行分型。

Flagellin gene typing of Campylobacter jejuni by restriction fragment length polymorphism analysis.

作者信息

Nachamkin I, Bohachick K, Patton C M

机构信息

Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia 19104-4283.

出版信息

J Clin Microbiol. 1993 Jun;31(6):1531-6. doi: 10.1128/jcm.31.6.1531-1536.1993.

Abstract

We developed and studied a molecular typing approach for Campylobacter spp. with restriction fragment length polymorphism (RFLP) analysis of the flagellin gene flaA in C. jejuni. Using polymerase chain reaction, we amplified the flaA gene from strains comprising different HL:O serotypes by using a primer set directed at the conserved 5' and 3' flaA gene sequence to generate a 1.7-kb amplicon. The amplicon was further digested with the restriction enzyme DdeI, and the fragments generated were analyzed by agarose gel electrophoresis. In 43 non-outbreak strains of six common HL serotypes (HL 1, 2, 4, 5, 9, and 36) in the United States, 18 RFLP patterns were observed. In U.S. outbreak strains previously studied by 10 other typing methods, flaA typing correlated with the HL serotype within each outbreak, and six additional flaA types were identified. Our results suggest that RFLP analysis of the flaA gene from Campylobacter spp. has sufficient discrimination to be useful as a practical typing method for clinical and epidemiologic investigations.

摘要

我们开发并研究了一种用于弯曲杆菌属的分子分型方法,该方法通过对空肠弯曲杆菌中鞭毛蛋白基因flaA进行限制性片段长度多态性(RFLP)分析来实现。利用聚合酶链反应,我们使用针对保守的flaA基因5'和3'序列的引物对,从包含不同HL:O血清型的菌株中扩增flaA基因,以产生一个1.7 kb的扩增子。扩增子再用限制性内切酶DdeI进行消化,然后通过琼脂糖凝胶电泳分析产生的片段。在美国的43株六种常见HL血清型(HL 1、2、4、5、9和36)的非暴发菌株中,观察到18种RFLP模式。在之前用其他10种分型方法研究过的美国暴发菌株中,flaA分型与每次暴发中的HL血清型相关,并且还鉴定出另外六种flaA类型。我们的结果表明,对弯曲杆菌属的flaA基因进行RFLP分析具有足够的区分能力,可作为临床和流行病学调查的实用分型方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9e05/265573/7c95059495e3/jcm00018-0145-a.jpg

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