Development of a Multiplex Recombinase Polymerase Amplification Coupled with Lateral Flow Dipsticks for the Simultaneous Rapid Detection of Salmonella spp., Salmonella Typhimurium and Salmonella Enteritidis

IF 3 3区农林科学 Q2 FOOD SCIENCE & TECHNOLOGY Food Quality and Safety Pub Date : 2023-11-23 DOI:10.1093/fqsafe/fyad059

Zeqiang Zhan, Shoukui He, Yan Cui, Jinzeng Yang, Xianming Shi

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Abstract

Salmonella spp. is one of the world-leading foodborne pathogens and its rapid detection is essential to ensure food safety. In this study, a visual method was established based on multiple recombinase polymerase amplification (RPA) coupled with lateral flow dipsticks (LFD) for the simultaneous detection of Salmonella spp., Salmonella Enteritidis (S. Enteritidis) and Salmonella Typhimurium (S. Typhimurium). The optimal volume and temperature for the multiplex RPA-LFD method were optimized as 25 μL and 38℃, respectively. The reaction process was completed within 25 min and the detection results were observed visually. The limit of detection (LOD) was 2.8×102, 5.9×102 and 7.6×102 CFU/mL for Salmonella spp., S. Enteritidis and S. Typhimurium, respectively. Meanwhile, the detection results of the established method showed no cross-reactivity between object Salmonella cells and other common foodborne bacteria, which was highly specific for Salmonella. More importantly, the developed method exhibited good performance in artificially contaminated chicken samples, and the LOD in artificially contaminated chicken samples was 2.8×103, 5.9×103 and 7.6×103 CFU/mL for Salmonella spp., S. Enteritidis and S. Typhimurium, respectively. Finally, the application of the multiple RPA-LFD method in retailed food samples displayed that this method was effectively and practically in the detection of Salmonella spp., S. Enteritidis and S. Typhimurium food. This multiplex RPA-LFD method provides a new sensitive and rapid alternative for the specific detection of Salmonella spp. and its important serovars in food.

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开发一种多重重组酶聚合酶扩增法与侧流浸渍棒相结合的方法，用于同时快速检测沙门氏菌属、鼠伤寒沙门氏菌和肠炎沙门氏菌

沙门氏菌是世界上主要的食源性致病菌之一，快速检测沙门氏菌对确保食品安全至关重要。本研究建立了一种基于多重重组酶聚合酶扩增（RPA）和侧流浸渍棒（LFD）的可视化方法，用于同时检测沙门氏菌属、肠炎沙门氏菌（S. Enteritidis）和鼠伤寒沙门氏菌（S. Typhimurium）。经优化，多重 RPA-LFD 法的最佳体积和温度分别为 25 μL 和 38℃。反应过程在 25 分钟内完成，检测结果通过肉眼观察。沙门氏菌属、肠炎沙门氏菌和鼠伤寒沙门氏菌的检出限（LOD）分别为 2.8×102、5.9×102 和 7.6×102 CFU/mL。同时，所建立方法的检测结果表明，目标沙门氏菌细胞与其他常见食源性细菌之间无交叉反应，对沙门氏菌具有高度特异性。更重要的是，所建立的方法在人工污染的鸡肉样品中表现出良好的性能，在人工污染的鸡肉样品中，沙门氏菌属、肠炎沙门氏菌和鼠伤寒沙门氏菌的LOD分别为2.8×103、5.9×103和7.6×103 CFU/mL。最后，多重 RPA-LFD 方法在零售食品样品中的应用表明，该方法在检测沙门氏菌属、肠炎沙门氏菌和鼠伤寒沙门氏菌食品方面是有效和实用的。这种多重 RPA-LFD 方法为特异性检测食品中的沙门氏菌及其重要血清型提供了一种新的灵敏而快速的选择。

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来源期刊

Food Quality and Safety FOOD SCIENCE & TECHNOLOGY-

CiteScore

7.20

自引率

1.80%

发文量

审稿时长

5 weeks

期刊介绍： Food quality and safety are the main targets of investigation in food production. Therefore, reliable paths to detect, identify, quantify, characterize and monitor quality and safety issues occurring in food are of great interest. Food Quality and Safety is an open access, international, peer-reviewed journal providing a platform to highlight emerging and innovative science and technology in the agro-food field, publishing up-to-date research in the areas of food quality and safety, food nutrition and human health. It promotes food and health equity which will consequently promote public health and combat diseases. The journal is an effective channel of communication between food scientists, nutritionists, public health professionals, food producers, food marketers, policy makers, governmental and non-governmental agencies, and others concerned with the food safety, nutrition and public health dimensions. The journal accepts original research articles, review papers, technical reports, case studies, conference reports, and book reviews articles.