Rapid Detection of Salmonella and Listeria monocytogenes in Fresh Beef Using Loop-Mediated Isothermal Amplification.

IF 1.9 2区农林科学 Q3 FOOD SCIENCE & TECHNOLOGY

Foodborne pathogens and disease Pub Date : 2025-04-28 DOI:10.1089/fpd.2025.0018

Xiaoran Wang, Kaixiang Jia, Jinrong Ran, Yangyang Du, Xiaoying Yu, Xuefeng Cao, Lianci Peng, Weifeng Chen, Rendong Fang, Zhiwei Li

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Abstract

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.

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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。

本研究建立并优化了环介导等温扩增（LAMP）快速检测新鲜牛肉中沙门氏菌和单核增生乳杆菌的方法。利用LAMP引物分别靶向沙门氏菌的invA基因和单核增生乳杆菌的hly基因。对不同病原菌的反应温度、dNTP浓度、Mg2+浓度、引物比例、Bst DNA聚合酶用量等条件进行优化。两种病原菌在最佳温度、Mg2+浓度和Bst DNA聚合酶需求方面存在差异，这凸显了病原菌特异性优化的重要性。优化后的方法灵敏度较高，沙门氏菌和单核增生乳杆菌的检出限分别为120 fg/μL和130 fg/μL，检测时间分别为40 min和60 min。特异性测试证实，这两种分析方法对新鲜牛肉样品中的目标病原体具有高度特异性，没有观察到交叉反应。添加羟基酚蓝使阳性结果的简单视觉检测，消除了对专用设备的需要。开发的LAMP检测方法可以快速、敏感和特异性地检测这些重要的食源性病原体。

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来源期刊

Foodborne pathogens and disease 医学-食品科技

CiteScore

5.30

自引率

3.60%

发文量

审稿时长

1 months

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.