一种基于铕纳米粒子的快速灵敏侧向流免疫分析与重组酶聚合酶扩增相结合的方法，用于同时检测三种食源性致病菌。

A Rapid and Sensitive Europium Nanoparticle-Based Lateral Flow Immunoassay Combined with Recombinase Polymerase Amplification for Simultaneous Detection of Three Food-Borne Pathogens.

机构信息

Zhejiang Provincial Key Laboratory of Biometrology and Inspection & Quarantine, China Jiliang University, Hangzhou 310018, China.

Department of Plant Biology, Uppsala BioCenter, Linnean Centre for Plant Biology, EuSwedish University of Agricultural Science (SLU), P.O. Box 7080, SE-75007 Uppsala, Sweden.

出版信息

Int J Environ Res Public Health. 2021 Apr 26;18(9):4574. doi: 10.3390/ijerph18094574.

DOI:10.3390/ijerph18094574

PMID:33925871

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8123443/

Abstract

Food-borne pathogens have become an important public threat to human health. There are many kinds of pathogenic bacteria in food consumed daily. A rapid and sensitive testing method for multiple food-borne pathogens is essential. Europium nanoparticles (EuNPs) are used as fluorescent probes in lateral flow immunoassays (LFIAs) to improve sensitivity. Here, recombinase polymerase amplification (RPA) combined with fluorescent LFIA was established for the simultaneous and quantitative detection of , , and . In this work, the entire experimental process could be completed in 20 min at 37 °C. The limits of detection (LODs) of EuNP-based LFIA-RPA were 9.0 colony-forming units (CFU)/mL for , 7.0 CFU/mL for , and 4.0 CFU/mL for . No cross-reaction could be observed in 22 bacterial strains. The fluorescent LFIA-RPA assay exhibits high sensitivity and good specificity. Moreover, the average recovery of the three food-borne pathogens spiked in food samples was 90.9-114.2%. The experiments indicate the accuracy and reliability of the multiple fluorescent test strips. Our developed EuNP-based LFIA-RPA assay is a promising analytical tool for the rapid and simultaneous detection of multiple low concentrations of food-borne pathogens.

摘要

食源性病原体已成为危害人类健康的重要公共卫生威胁。日常食用的食物中存在多种致病菌。因此，建立一种快速、灵敏的多重食源性致病菌检测方法至关重要。本研究将上转换纳米粒子（EuNPs）作为侧向流动免疫分析（LFIAs）中的荧光探针，以提高其灵敏度。本研究建立了重组酶聚合酶扩增（RPA）与荧光 LFIAs 相结合的方法，用于同时定量检测、、。在本工作中，整个实验过程可在 37°C 下 20 min 内完成。基于 EuNP 的 LFIA-RPA 的检测限（LOD）分别为 9.0 菌落形成单位（CFU）/mL 、7.0 CFU/mL 和 4.0 CFU/mL。在 22 种细菌菌株中未观察到交叉反应。荧光 LFIA-RPA 检测法具有较高的灵敏度和良好的特异性。此外，三种食源性致病菌在食物样本中的平均回收率为 90.9-114.2%。实验表明，该多重荧光检测试纸具有较高的准确性和可靠性。本研究开发的基于 EuNP 的 LFIA-RPA 检测方法是一种用于快速、同时检测多种低浓度食源性致病菌的有前途的分析工具。