侧向流动型重组酶聚合酶扩增技术检测食品中大肠埃希氏菌O157  

The Detection Method for Escherichia Coli O157 Based on Lateral Flow-Based Recombinase Polymerase Amplification

作　　者：兰全学 陈佳平 杨慧[1] 甘莉萍[1] 杜文旗 刘渠[1] 金玉娟[1] LAN Quanxue;CHEN Jiaping;YANG Hui;GAN Liping;DU Wenqi;LIU Qu;JIN Yujuan(Shenzhen Longgang District Center for Disease Control and Prevention,Shenzhen 518172,China;Shenzhen Academy of Metrology&Quality Inspection,Shenzhen 518109,China)

机构地区：[1]深圳市龙岗区疾病预防控制中心,广东深圳518172 [2]深圳市计量质量检测研究院,广东深圳518109

出　　处：《食品科技》2021年第1期302-307,共6页Food Science and Technology

基　　金：深圳市科创委项目(JSGG20160606144217004)。

摘　　要：研究内容基于侧向流动型重组酶聚合酶扩增(Lateral flow-based recombinase polymerase amplification,LF-RPA)技术原理,建立大肠埃希氏菌O157(Escherichia coli O157)的LF-RPA快速检测方法。应用该方法对3株大肠埃希氏菌O157和27株非大肠埃希氏菌O157进行检测,结果表明该方法具有良好的特异性。分别以10倍浓度梯度稀释插入靶基因序列的质粒pUC57-rfbE和大肠埃希氏菌O157菌液DNA作为模板进行LF-RPA检测。结果表明LF-RPA方法可检出1.0×10^(2)拷贝/μL的pUC57-rfbE质粒和1.2×10^(4) cfu/mL的大肠埃希氏菌O157菌液。以大肠埃希氏菌O157污染食品样品,增菌4 h后,大肠埃希氏菌O157初始浓度为1.2×10^(1) cfu/mL的食品增菌液可以被LF-RPA检出阳性信号。该研究建立的方法可快速、准确、灵敏地检测食品中大肠埃希氏菌O157,无需专业设备,在食品生产企业和基层检测机构具有一定的应用前景。This study developed a rapid detection method for Escherichia coli O157 based on lateral flow-based recombinase polymerase amplification (LF-RPA).According to testing 3 Escherichia coli O157 and 27 non-Escherichia coli O157 strains by using LF-RPA,the developed method in this research can detect Escherichia coli O157 specifically.To carry out LF-RPA assay with the templates of tenfold gradient diluted pUC57-rfbE plasmids;moreover,LF-RPA assay was carried out by using the templates of extracted DNA from tenfold gradient diluted Escherichia coli O157 liquid.The sensitivity of LF-RPA can reach 1.0×10^(2) copies/μL and 1.2×10^(4) cfu/mL,respectively.The food samples were inoculated with Escherichia coli O157,positive signals can be obtained from the food samples inoculated with 1.2×10^(1) cfu/mL Escherichia coli O157 after 4 h enrichment.This study developed a testing method for Escherichia coli O157,this method can detect Escherichia coli O157 rapidly,specifically and sensitively without professional equipment.So Escherichia coli O157 LF-RPA can be used for point-of-care detection of Escherichia coli O157 in food production enterprises and grass-roots testing institutions.

关 键 词：重组酶聚合酶扩增 侧向流动免疫分析 大肠埃希氏菌O157 rfbE基因 

分 类 号：TS207.4[轻工技术与工程—食品科学]