Development and Evaluation of a Multiplex Fluorescence PCR for Salmonella Virulence Genes Analysis

IF 0.5 4区医学 Q4 MICROBIOLOGY

Jundishapur Journal of Microbiology Pub Date : 2024-05-12 DOI:10.5812/jjm-144579

Peiyan He, Yong Yan, Guoying Zhu, Ze Zhu, Zhongwen Chen

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Abstract

Background: Salmonellosis, a disease caused by Salmonella, is a significant public health concern and economic burden worldwide. The ability of various Salmonella serovars to cause disease is closely linked to the virulence genes they possess. Objectives: The aim of this study was to develop a multiplex fluorescence PCR for detecting ten major virulence genes (ssaR, spvC, pefA, sipA, fimA, sifA, sopE2, sopB, prgH, and stn) in Salmonella. Methods: Primer pairs specific to ten target virulence genes were designed using Primer Premier 5.0 and distributed across two reaction tubes. The multiplex fluorescence PCR was optimized by adjusting one factor at a time. Results: A total of sixty Salmonella strains were analyzed using the newly developed multiplex fluorescence PCR. All strains contained seven or more of the tested virulence genes. The positive rates of virulence genes ssaR, sipA, sopE2, sopB, prgH, and stn were high, each at 100%. The positive rate of sifA was also relatively high at 81.67%. However, the positive rates of spvC at 5% and pefA at 3.33% were relatively low. Conclusions: The newly developed multiplex fluorescence PCR provides a straightforward, cost-effective, and high-throughput solution for detecting virulence genes in Salmonella. It has the potential to become a routine method for analyzing Salmonella virulence genes.

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用于沙门氏菌病毒基因分析的多重荧光 PCR 的开发与评估

背景：沙门氏菌病是由沙门氏菌引起的一种疾病，是全球重大的公共卫生问题和经济负担。各种沙门氏菌血清型的致病能力与其所拥有的毒力基因密切相关。研究目的本研究旨在开发一种多重荧光 PCR，用于检测沙门氏菌中的 10 个主要毒力基因（ssaR、spvC、pefA、sipA、fimA、sifA、sopE2、sopB、prgH 和 stn）。方法：使用 Primer Premier 5.0 设计了特异于 10 个目标毒力基因的引物对，并将其分布在两个反应管中。通过每次调整一个因子来优化多重荧光 PCR。结果使用新开发的多重荧光 PCR 分析了六十株沙门氏菌。所有菌株都含有 7 个或更多被测毒力基因。毒力基因 ssaR、sipA、sopE2、sopB、prgH 和 stn 的阳性率很高，均为 100%。sifA 的阳性率也相对较高，为 81.67%。不过，spvC 的阳性率为 5%，pefA 的阳性率为 3.33%，相对较低。结论新开发的多重荧光 PCR 为检测沙门氏菌毒力基因提供了一种简单、经济、高通量的解决方案。它有望成为分析沙门氏菌毒力基因的常规方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Jundishapur Journal of Microbiology MICROBIOLOGY-

CiteScore

1.30

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： Jundishapur Journal of Microbiology, (JJM) is the official scientific Monthly publication of Ahvaz Jundishapur University of Medical Sciences. JJM is dedicated to the publication of manuscripts on topics concerning all aspects of microbiology. The topics include medical, veterinary and environmental microbiology, molecular investigations and infectious diseases. Aspects of immunology and epidemiology of infectious diseases are also considered.