一种用于鉴定沙门氏菌亚种的新型多重 PCR 检测方法。

A novel multiplex PCR assay for Salmonella subspecies identification.

机构信息

Division of Animal Life Science, Institute of Symbiotic Science and Technology, Tokyo University of Agriculture and Technology, Tokyo, Japan.

出版信息

J Appl Microbiol. 2009 Sep;107(3):805-11. doi: 10.1111/j.1365-2672.2009.04263.x. Epub 2009 Mar 31.

DOI:10.1111/j.1365-2672.2009.04263.x

PMID:19486419

Abstract

AIM

To develop a novel multiplex polymerase chain reaction (PCR) assay with six primer pairs for Salmonella subspecies identification.

METHODS AND RESULTS

Five primer pairs were chosen to detect the genes (fljB, mdcA, gatD, stn and STM4057) responsible for several phenotypic traits or encoding (sub) species-specific regions. A primer pair for invA was added to simultaneously detect Salmonella. The combination of these primer pairs was expected to give unique results to all subspecies, including Salmonella bongori. The multiplex PCR assay was optimized and evaluated with 53 Salmonella strains representing all S. enterica subspecies, S. bongori and five non-Salmonella strains. The multiplex PCR assay revealed that the genotypes were well correlated with the phenotypes in the Salmonella strains tested. The unique band patterns to their subspecies were generated from 94.3% (50/53) of the Salmonella strains, and no product from other strains by the multiplex PCR assay.

CONCLUSIONS

The multiplex PCR assay we developed was found to be a rapid, specific and easy to perform method compared with traditional biochemical tests for Salmonella subspecies identification, especially for rapid screening of large numbers of samples.

SIGNIFICANCE AND IMPACT OF THE STUDY

The assay will be useful for characterizing Salmonella isolates from reptiles, which belong to various subspecies, and therefore add to the scientific understanding of reptile-associated Salmonellosis.

摘要

目的

开发一种新的多重聚合酶链反应（PCR）检测方法，使用 6 对引物对沙门氏菌亚种进行鉴定。

方法和结果

选择了 5 对引物来检测负责多个表型特征的基因（fljB、mdcA、gatD、stn 和 STM4057）或编码（亚）种特异性区域。添加了一对引物 invA 来同时检测沙门氏菌。这些引物对的组合预计将为所有亚种，包括沙门氏菌 bongori 提供独特的结果。优化并评估了多重 PCR 检测方法，使用 53 株代表所有肠沙门氏菌亚种、沙门氏菌 bongori 和 5 株非沙门氏菌的菌株进行检测。多重 PCR 检测显示，基因型与测试的沙门氏菌菌株的表型高度相关。来自 94.3%（50/53）的沙门氏菌菌株的亚种独特的带型是通过多重 PCR 检测产生的，而其他菌株则没有产物。