一种针对 gyrB 的 PCR 方法，可快速鉴定沙门氏菌。

A gyrB-targeted PCR for rapid identification of Salmonella.

机构信息

State Key Laboratory of Soil and Sustainable Agriculture, Institute of Soil Science, Chinese Academy of Sciences, Beijing East Road, 71, Nanjing 210008, People's Republic of China.

出版信息

Curr Microbiol. 2011 Nov;63(5):477-83. doi: 10.1007/s00284-011-0007-1. Epub 2011 Sep 3.

DOI:10.1007/s00284-011-0007-1

PMID:21892633

Abstract

Salmonella causes the majority of infections in humans and homeothermic animals. This article describes a specific polymerase chain reaction (PCR) method developed for a rapid identification of Salmonella. A gyrB-targeted species-specific primer pair, S-P-for (5'-GGT GGT TTC CGT AAA AGT A-3') and S-P-rev (5'-GAA TCG CCT GGT TCT TGC-3'), was successfully designed. PCR with all the Salmonella strains produced a 366- bp DNA fragment that was absent from all the non-Salmonella strains tested. The detection limit of the PCR was 0.01 ng with genomic DNA or 3.2 cells per assay. Good specificity was also demonstrated by fecal samples, from which only the gyrB gene of Salmonella was amplified. Using the culture-PCR method, 27 isolates on Salmonella-Shigella (SS) medium were rapidly identified as Salmonella, which was confirmed by the sequencing of the gyrB gene.

摘要

沙门氏菌引起人类和恒温动物的大多数感染。本文描述了一种快速鉴定沙门氏菌的特定聚合酶链反应（PCR）方法。成功设计了一对针对gyrB 的种特异性引物，S-P-for（5'-GGT GGT TTC CGT AAA AGT A-3'）和 S-P-rev（5'-GAA TCG CCT GGT TCT TGC-3'）。所有沙门氏菌菌株的 PCR 均产生了 366-bp 的 DNA 片段，而所有测试的非沙门氏菌菌株均未产生该片段。PCR 的检测限为 0.01ng 基因组 DNA 或每个检测 3.2 个细胞。粪便样本也表现出良好的特异性，仅扩增出沙门氏菌的 gyrB 基因。使用培养-PCR 方法，快速鉴定出 SS 培养基上的 27 个分离物为沙门氏菌，这通过 gyrB 基因的测序得到了确认。