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基于cadF毒力基因及其产物鉴定空肠弯曲菌和结肠弯曲菌这两种肠道病原体。

Identification of the enteropathogens Campylobacter jejuni and Campylobacter coli based on the cadF virulence gene and its product.

作者信息

Konkel M E, Gray S A, Kim B J, Garvis S G, Yoon J

机构信息

Department of Microbiology, Washington State University, Pullman, Washington 99164-4233, USA.

出版信息

J Clin Microbiol. 1999 Mar;37(3):510-7. doi: 10.1128/JCM.37.3.510-517.1999.

Abstract

Campylobacter jejuni and Campylobacter coli are common causes of gastroenteritis in humans. Infection with C. jejuni or C. coli is commonly acquired by eating undercooked chicken. The goal of this study was to develop specific detection assays for C. jejuni and C. coli isolates based on the cadF virulence gene and its product. The cadF gene from C. jejuni and C. coli encodes a 37-kDa outer membrane protein that promotes the binding of these pathogens to intestinal epithelial cells. A fragment of approximately 400 bp was amplified from 38 of 40 (95%) C. jejuni isolates and 5 of 6 (83.3%) C. coli isolates with primers designed to amplify an internal fragment of the cadF gene. PCR was then used to amplify Campylobacter DNA from store-bought chickens. A 400-bp band was amplified from 26 of the 27 chicken carcasses tested by the PCR-based assay. The CadF protein was detected in every C. jejuni and C. coli isolate tested, as judged by immunoblot analysis with a rabbit anti-C. jejuni 37-kDa serum. In addition, methanol-fixed samples of whole-cell C. jejuni and C. coli were detected with the rabbit anti-37-kDa serum by using an indirect-immunofluorescence microscopy assay. These findings indicate that the cadF gene and its product are conserved among C. jejuni and C. coli isolates and that a PCR assay based on the cadF gene may be useful for the detection of Campylobacter organisms in food products.

摘要

空肠弯曲菌和结肠弯曲菌是人类肠胃炎的常见病因。感染空肠弯曲菌或结肠弯曲菌通常是由于食用未煮熟的鸡肉所致。本研究的目的是基于cadF毒力基因及其产物开发针对空肠弯曲菌和结肠弯曲菌分离株的特异性检测方法。空肠弯曲菌和结肠弯曲菌的cadF基因编码一种37 kDa的外膜蛋白,该蛋白可促进这些病原体与肠道上皮细胞的结合。用设计用于扩增cadF基因内部片段的引物,从40株空肠弯曲菌分离株中的38株(95%)和6株结肠弯曲菌分离株中的5株(83.3%)扩增出了一段约400 bp的片段。然后用PCR从市售鸡肉中扩增弯曲菌DNA。基于PCR的检测方法在检测 的27个鸡 carcasses中的26个中扩增出了一条400 bp的条带。通过用兔抗空肠弯曲菌37 kDa血清进行免疫印迹分析判断,在每一株检测的空肠弯曲菌和结肠弯曲菌分离株中都检测到了CadF蛋白。此外,通过间接免疫荧光显微镜检测法,用兔抗37 kDa血清检测了全细胞空肠弯曲菌和结肠弯曲菌的甲醇固定样本。这些结果表明,cadF基因及其产物在空肠弯曲菌和结肠弯曲菌分离株中是保守 的,基于cadF基因的PCR检测方法可能有助于检测食品中的弯曲菌。 (注:原文中“鸡carcasses”处“检测的27个鸡carcasses中的26个”中“检测的”前少了“对”字,翻译时根据上下文补充完整了句子结构和意思。)

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