Xiaoran Wang, Kaixiang Jia, Jinrong Ran, Yangyang Du, Xiaoying Yu, Xuefeng Cao, Lianci Peng, Weifeng Chen, Rendong Fang, Zhiwei Li
{"title":"环介导等温扩增快速检测新鲜牛肉中沙门氏菌和单核增生李斯特菌。","authors":"Xiaoran Wang, Kaixiang Jia, Jinrong Ran, Yangyang Du, Xiaoying Yu, Xuefeng Cao, Lianci Peng, Weifeng Chen, Rendong Fang, Zhiwei Li","doi":"10.1089/fpd.2025.0018","DOIUrl":null,"url":null,"abstract":"<p><p>This study developed and optimized loop-mediated isothermal amplification (LAMP) assays for rapid detection of <i>Salmonella</i> and <i>L. monocytogenes</i> in fresh beef. LAMP primers targeting the <i>invA</i> gene of <i>Salmonella</i> and <i>hly</i> gene of <i>L. monocytogenes</i> were used. Reaction conditions including temperature, dNTP concentration, Mg<sup>2+</sup> concentration, primer ratio, and Bst DNA polymerase amount were optimized for each pathogen. Differences were observed between the two pathogens in optimal temperature, Mg<sup>2+</sup> concentration, and Bst DNA polymerase requirements, highlighting the importance of pathogen-specific optimization. The optimized assays demonstrated high sensitivity with detection limits of 120 fg/μL for <i>Salmonella</i> and 130 fg/μL for <i>L. monocytogenes</i>, achieving detection within 40 and 60 min, respectively. Specificity tests confirmed both assays were highly specific for their target pathogens in fresh beef samples with no cross-reactivity observed. Addition of hydroxynaphthol blue enabled simple visual detection of positive results, eliminating the need for specialized equipment. The developed LAMP assays offer rapid, sensitive, and specific detection of these important foodborne pathogens.</p>","PeriodicalId":12333,"journal":{"name":"Foodborne pathogens and disease","volume":" ","pages":""},"PeriodicalIF":1.9000,"publicationDate":"2025-04-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Rapid Detection of <i>Salmonella</i> and <i>Listeria monocytogenes</i> in Fresh Beef Using Loop-Mediated Isothermal Amplification.\",\"authors\":\"Xiaoran Wang, Kaixiang Jia, Jinrong Ran, Yangyang Du, Xiaoying Yu, Xuefeng Cao, Lianci Peng, Weifeng Chen, Rendong Fang, Zhiwei Li\",\"doi\":\"10.1089/fpd.2025.0018\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>This study developed and optimized loop-mediated isothermal amplification (LAMP) assays for rapid detection of <i>Salmonella</i> and <i>L. monocytogenes</i> in fresh beef. LAMP primers targeting the <i>invA</i> gene of <i>Salmonella</i> and <i>hly</i> gene of <i>L. monocytogenes</i> were used. Reaction conditions including temperature, dNTP concentration, Mg<sup>2+</sup> concentration, primer ratio, and Bst DNA polymerase amount were optimized for each pathogen. Differences were observed between the two pathogens in optimal temperature, Mg<sup>2+</sup> concentration, and Bst DNA polymerase requirements, highlighting the importance of pathogen-specific optimization. The optimized assays demonstrated high sensitivity with detection limits of 120 fg/μL for <i>Salmonella</i> and 130 fg/μL for <i>L. monocytogenes</i>, achieving detection within 40 and 60 min, respectively. Specificity tests confirmed both assays were highly specific for their target pathogens in fresh beef samples with no cross-reactivity observed. Addition of hydroxynaphthol blue enabled simple visual detection of positive results, eliminating the need for specialized equipment. The developed LAMP assays offer rapid, sensitive, and specific detection of these important foodborne pathogens.</p>\",\"PeriodicalId\":12333,\"journal\":{\"name\":\"Foodborne pathogens and disease\",\"volume\":\" \",\"pages\":\"\"},\"PeriodicalIF\":1.9000,\"publicationDate\":\"2025-04-28\",\"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.2025.0018\",\"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}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Foodborne pathogens and disease","FirstCategoryId":"97","ListUrlMain":"https://doi.org/10.1089/fpd.2025.0018","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}
Rapid Detection of Salmonella and Listeria monocytogenes in Fresh Beef Using Loop-Mediated Isothermal Amplification.
This study developed and optimized loop-mediated isothermal amplification (LAMP) assays for rapid detection of Salmonella and L. monocytogenes in fresh beef. LAMP primers targeting the invA gene of Salmonella and hly gene of L. monocytogenes were used. Reaction conditions including temperature, dNTP concentration, Mg2+ concentration, primer ratio, and Bst DNA polymerase amount were optimized for each pathogen. Differences were observed between the two pathogens in optimal temperature, Mg2+ concentration, and Bst DNA polymerase requirements, highlighting the importance of pathogen-specific optimization. The optimized assays demonstrated high sensitivity with detection limits of 120 fg/μL for Salmonella and 130 fg/μL for L. monocytogenes, achieving detection within 40 and 60 min, respectively. Specificity tests confirmed both assays were highly specific for their target pathogens in fresh beef samples with no cross-reactivity observed. Addition of hydroxynaphthol blue enabled simple visual detection of positive results, eliminating the need for specialized equipment. The developed LAMP assays offer rapid, sensitive, and specific detection of these important foodborne pathogens.
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.