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基于重组酶聚合酶扩增与侧向流试纸条(RPA-LFD)的肉中三种食源性致病菌的灵敏快速检测。

Sensitive and rapid detection of three foodborne pathogens in meat by recombinase polymerase amplification with lateral flow dipstick (RPA-LFD).

机构信息

Key Laboratory for Food Microbial Technology of Zhejiang Province, School of Food Science and Biotechnology, Zhejiang Gongshang niversity, No. 18, Xuezheng Str, Hangzhou 310018, China.

Assure Tech. (Hangzhou) Co., Ltd, Building 4, No. 1418-50, Moganshan Road, Gongshu District, Hangzhou 310011, China.

出版信息

Int J Food Microbiol. 2024 Sep 16;422:110822. doi: 10.1016/j.ijfoodmicro.2024.110822. Epub 2024 Jul 3.

Abstract

Foodborne illnesses, caused by harmful microorganisms in food, are a significant global health issue. Current methods for identifying these pathogens are both labor-intensive and time-consuming. In this research, we devised a swift and precise detection technique using recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD) for three foodborne pathogens found in meat. By employing a dedicated detection device, RPA-LFD allows for the rapid analysis of DNA from Escherichia coli O157 (E. coli O157), Salmonella, and Shigella-pathogens that are prohibited in food. The detection thresholds for E. coli O157, Salmonella, and Shigella are 0.168 fg/μl (1.04 CFU/ml), 0.72 fg/μl (27.49 CFU/ml), and 1.25 fg/μl (48.84 CFU/ml), respectively. This method provides a short detection window, operates at low temperatures, follows simple procedures, and exhibits high sensitivity. Our study establishes the RPA-LFD method for simultaneously identifying the nucleic acid of three foodborne pathogens, offering an efficient solution for quickly identifying multiple contaminants.

摘要

食源性疾病是由食物中的有害微生物引起的,是一个严重的全球健康问题。目前识别这些病原体的方法既费时又费力。在这项研究中,我们设计了一种快速而精确的检测技术,使用重组酶聚合酶扩增结合横向流动纸条(RPA-LFD)检测肉类中的三种食源性病原体。通过使用专用的检测设备,RPA-LFD 可以快速分析大肠杆菌 O157(E. coli O157)、沙门氏菌和志贺氏菌的 DNA,这些病原体在食品中是被禁止的。E. coli O157、沙门氏菌和志贺氏菌的检测阈值分别为 0.168 fg/μl(1.04 CFU/ml)、0.72 fg/μl(27.49 CFU/ml)和 1.25 fg/μl(48.84 CFU/ml)。该方法具有检测窗口短、低温操作、简单的操作步骤和高灵敏度的特点。我们的研究建立了 RPA-LFD 方法,用于同时鉴定三种食源性病原体的核酸,为快速鉴定多种污染物提供了有效的解决方案。

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