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基于一步聚合酶链反应的弓形杆菌属分型

One-step polymerase chain reaction-based typing of Arcobacter species.

作者信息

Kabeya Hidenori, Kobayashi Yoko, Maruyama Soichi, Mikami Takeshi

机构信息

Laboratory of Veterinary Public Health, Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, Kameino, Fujisawa, Kanagawa 252-8510, Japan.

出版信息

Int J Food Microbiol. 2003 Mar 15;81(2):163-8. doi: 10.1016/s0168-1605(02)00197-6.

Abstract

A species-specific PCR assay was developed for the identification of the Arcobacter species, Arcobacter butzleri, Arcobacter cryaerophilus 1A and 1B, and Arcobacter skirrowii. The primers, which amplify the most variable areas of the 23S rRNA gene, were designed to perform species-specific identification by one-step PCR. The DNA sequence of the region from A. cryaerophilus 1B was determined, and the specific pimer for the species was designed. By using one-step PCR containing the mixed primers N.butz, N.c1.A, N.c.1B, and N.ski, species-specific amplifications were detected from the reference strains of A. butzleri, A. cryaerophilus 1A, 1B, and A. skirrowii, respectively. Primers designed in this study were also evaluated on 10 of Japanese field isolates, and all species were identified. This simple one-step PCR assay was found to be a powerful tool for the survey of Arcobacter infection.

摘要

开发了一种种特异性聚合酶链反应(PCR)检测方法,用于鉴定嗜低温栖热菌属细菌、布氏嗜低温栖热菌、嗜低温栖热菌1A和1B以及斯氏嗜低温栖热菌。这些引物可扩增23S核糖体RNA(rRNA)基因的最可变区域,旨在通过一步PCR进行种特异性鉴定。测定了嗜低温栖热菌1B该区域的DNA序列,并设计了该物种的特异性引物。通过使用含有混合引物N.butz、N.c1.A、N.c.1B和N.ski的一步PCR,分别从布氏嗜低温栖热菌、嗜低温栖热菌1A、1B和斯氏嗜低温栖热菌的参考菌株中检测到种特异性扩增。本研究设计的引物也在10株日本野外分离株上进行了评估,所有菌种均得以鉴定。结果发现,这种简单的一步PCR检测方法是调查嗜低温栖热菌感染的有力工具。

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