{"title":"Development and Application of Four Foodborne Pathogens by TaqMan Multiplex Real-Time PCR.","authors":"Yinlei Xue, Shengfang He, Meng Li, Yuanhao Qiu","doi":"10.1089/fpd.2023.0134","DOIUrl":null,"url":null,"abstract":"<p><p>A TaqMan multiplex real-time PCR (mRT-PCR) was developed to detect simultaneously <i>Salmonella</i> spp., <i>Escherichia coli</i> O157, <i>Staphylococcus aureus</i>, and <i>Listeria monocytogene</i>s in food samples. The method involves four sets of primers and probes tailored to the unique DNA sequences found in the <i>invA</i>, <i>nuc</i>, <i>rfbE</i>, and <i>hly</i> genes of each pathogen. The generated standard curves, correlating gene copy numbers with Ct values, demonstrated high accuracy (<i>R</i><sup>2</sup> > 0.99) and efficiency (92%-104%). Meanwhile, the limit of detection was 10<sup>0</sup> CFU/mL for the four target bacteria in artificially contaminated food samples after 6-8 h of enrichment. The assay's effectiveness was further verified by testing 80 naturally contaminated food samples, showing results largely in agreement with traditional culture methods. Overall, this newly developed TaqMan mRT-PCR, inclusive of a pre-enrichment step, proves to be a dependable and effective tool for detecting single or multiple pathogens in diverse food items, offering significant potential for <i>in vitro</i> diagnostics.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":""},"PeriodicalIF":1.9000,"publicationDate":"2024-04-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Foodborne pathogens and disease","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1089/fpd.2023.0134","RegionNum":2,"RegionCategory":"农林科学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"FOOD SCIENCE & TECHNOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
A TaqMan multiplex real-time PCR (mRT-PCR) was developed to detect simultaneously Salmonella spp., Escherichia coli O157, Staphylococcus aureus, and Listeria monocytogenes in food samples. The method involves four sets of primers and probes tailored to the unique DNA sequences found in the invA, nuc, rfbE, and hly genes of each pathogen. The generated standard curves, correlating gene copy numbers with Ct values, demonstrated high accuracy (R2 > 0.99) and efficiency (92%-104%). Meanwhile, the limit of detection was 100 CFU/mL for the four target bacteria in artificially contaminated food samples after 6-8 h of enrichment. The assay's effectiveness was further verified by testing 80 naturally contaminated food samples, showing results largely in agreement with traditional culture methods. Overall, this newly developed TaqMan mRT-PCR, inclusive of a pre-enrichment step, proves to be a dependable and effective tool for detecting single or multiple pathogens in diverse food items, offering significant potential for in vitro diagnostics.
期刊介绍:
Foodborne Pathogens and Disease is one of the most inclusive scientific publications on the many disciplines that contribute to food safety. Spanning an array of issues from "farm-to-fork," the Journal bridges the gap between science and policy to reduce the burden of foodborne illness worldwide.
Foodborne Pathogens and Disease coverage includes:
Agroterrorism
Safety of organically grown and genetically modified foods
Emerging pathogens
Emergence of drug resistance
Methods and technology for rapid and accurate detection
Strategies to destroy or control foodborne pathogens
Novel strategies for the prevention and control of plant and animal diseases that impact food safety
Biosecurity issues and the implications of new regulatory guidelines
Impact of changing lifestyles and consumer demands on food safety.