基于侧向流免疫的单增李斯特氏菌检测方法的建立  

Establishment of A Detection Method for Listeria monocytogenes Based on Lateral Flow Immunity

作　　者：郦娟 陈琼 刘萍 褚冲 戴小芳 辛博 高家昊 杨永 LI Juan;CHEN Qiong;LIU Ping;CHU Chong;DAI Xiaofang;XIN Bo;GAO Jiahao;YANG Yong(Wuhan Institute for Food and Cosmetic Control,Key Laboratory of National Market Supervision(Edible Oil Quality and Safety),Wuhan 430040,China)

机构地区：[1]武汉食品化妆品检验所,国家市场监管重点实验室(食用油质量与安全),湖北武汉430040

出　　处：《食品科技》2024年第3期344-350,共7页Food Science and Technology

基　　金：国家市场监督管理总局科技项目(2020MK131);湖北省市场监督管理局技术保障和科技计划项目(Hbscjg-JS2021007)。

摘　　要：传统微生物方法以及实时荧光聚合酶链式反应(Polymerase chain reaction,PCR)方法都不能完全满足重大活动食品安全保障工作中现场检测的要求,因此研究将重组酶聚合酶等温扩增(Recombinant polymerase isothermal amplification,RPA)与侧向流免疫试纸(Lateral flow immunoassay,LFS)技术结合用于检测单增李斯特氏菌。RPA-LFS方法在纯菌、人工污染熟肉制品和新鲜水果中的检出限分别为3.0×10^(1)、3.0×10^(1)CFU/g和3.0×10^(2)CFU/g。结合叠氮溴化丙啶(Propidium monoazide,PMA)处理,建立了检测活菌的PMA-RPA-LFS检测方法,该方法可以抑制10^(4)CFU/mL浓度水平的死菌的扩增。将2种方法和GB 4789.10—2016用于人工污染和实际样品的检测,结果发现PMARPA-LFS可减少死菌带来的假阳性。建立的方法分别适用于未经或经热处理的食品。该研究为不同类型食品提供不同的检测方法选择,同时为重大活动食品安全保障工作中致病菌的检测提供了新思路。Traditional microbial methods and real-time fluorescence polymerase chain reaction(PCR)methods could not fully meet the requirements of on-site testing in food safety assurance work for major events.Recombinant polymerase isothermal amplification(RPA)and lateral flow immunoassay(LFS)technology were combined together to detect Listeria monocytogenes in this study.The detection limit of the RPA-LFS in the pure bacteria,artificially contaminated meat products and artificially contaminated fresh fruits were 3.0×10^(1) CFU/mL,3.0×10^(1) CFU/g and 3.0×10^(2) CFU/g respectively.A detection method for live bacteria was established by combining with propidium monoazide(PMA)treatment,which could inhibit the amplification of dead bacteria at a concentration level of 10^(4) CFU/mL.The two methods and GB 4789.10—2016 were used for the detection of artificial contamination and actual samples,and it was found that PMA-RPA-LFS reduced false positives caused by dead bacteria.The established methods were applicable to food that has not been or has been heat treated.This study provided different choices for different types of food and new ideas for the detection of pathogenic bacteria in food safety assurance work during major events.

关 键 词：单增李斯特氏菌 叠氮溴化丙锭 侧向流免疫 重组酶聚合酶等温扩增 

分 类 号：TS207.4[轻工技术与工程—食品科学]