不对称PCR技术结合免疫层析法快速检测产志贺毒素大肠埃希氏菌

doi:10.7506/spkx1002-6630-20200113-129

食品科学 ›› 2021, Vol. 42 ›› Issue (2): 312-318.doi: 10.7506/spkx1002-6630-20200113-129

不对称PCR技术结合免疫层析法快速检测产志贺毒素大肠埃希氏菌

山珊，黄昭鸿，黄艳梅，刘道峰，刘成伟，黄运红，龙中儿

（1.江西师范大学生命科学学院，南昌市鄱阳湖湿地微生物资源开发与利用重点实验室，江西南昌 330022；2.江西业力医疗器械有限公司，江西南昌 330008；3.江西省疾病预防控制中心，江西省食源性疾病诊断溯源重点实验室，江西南昌 330029）

出版日期:2021-01-18 发布日期:2021-01-27
基金资助:
江西省卫健委科技计划项目（20201120）

Rapid Detection of Shiga Toxin-producing Escherichia coli by Immunochromatography Combined with Asymmetric Polymerase Chain Reaction

SHAN Shan, HUANG Zhaohong, HUANG Yanmei, LIU Daofeng, LIU Chengwei, HUANG Yunhong, LONG Zhong’er

(1. Nanchang Key Laboratory of Microbial Resources Exploitation & Utilization from Poyang Lake Wetland, College of Life Sciences, Jiangxi Normal University, Nanchang 330022, China; 2. Jiangxi Yeli Medical Device Co. Ltd., Nanchang 330008, China;3. Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Province Center for Disease Control and Prevention, Nanchang 330029, China)

Online:2021-01-18 Published:2021-01-27

摘要/Abstract

摘要： 产志贺毒素大肠埃希氏菌（Shiga toxin-producing Escherichia coli，STEC）可分泌志贺毒素，致病性强，建立简便快速的STEC检测方法对控制食源性疾病的发生和蔓延极为重要。使用生物素标记STEC的标志性毒力基因stx1与stx2的下游引物进行不对称聚合酶链式反应，获得生物素化的目标单链DNA与聚苯乙烯微球标记的目标基因的上游引物特异性结合形成结合物，结合物上的生物素因与试纸条检测线上链霉亲和素结合而使检测线显色。通过建立的方法可成功快速检测出携带不同毒力基因的STEC，同时利用该方法对23 株菌株进行stx基因的检测，结果显示该方法可准确检测到含有STEC标志性毒力基因stx的菌株，且表现出良好的特异性。

关键词: 不对称聚合酶链式反应；免疫层析技术；产志贺毒素大肠埃希氏菌；检测

Abstract: Shiga toxin-producing Escherichia coli (STEC) is strongly pathogenic. Establishing a simple and rapid method for STEC detection is of importance for controlling the occurrence and spread of foodborne outbreaks. In this paper, the biotinylated target single stranded DNA was obtained by asymmetric polymerase chain reaction (aPCR) with the biotinylated downstream primers of the typical virulence genes stx1 and stx2. The DNA amplification products were conjugated with the upstream primers which were labeled on polystyrene microspheres. The biotin from the conjugates could be captured by streptavidin sprayed on a lateral flow strip, forming a red line visible to the naked eye. STEC strains producing different types of Shiga toxin could be detected by this method. It displayed a high specificity and accuracy for detecting 23 strains.

Key words: asymmetric polymerase chain reaction; immunochromatography; Shiga toxin-producing Escherichia coli; detection

中图分类号:

山珊，黄昭鸿，黄艳梅，刘道峰，刘成伟，黄运红，龙中儿. 不对称PCR技术结合免疫层析法快速检测产志贺毒素大肠埃希氏菌[J]. 食品科学, 2021, 42(2): 312-318.

SHAN Shan, HUANG Zhaohong, HUANG Yanmei, LIU Daofeng, LIU Chengwei, HUANG Yunhong, LONG Zhong’er. Rapid Detection of Shiga Toxin-producing Escherichia coli by Immunochromatography Combined with Asymmetric Polymerase Chain Reaction[J]. FOOD SCIENCE, 2021, 42(2): 312-318.

不对称PCR技术结合免疫层析法快速检测产志贺毒素大肠埃希氏菌

Rapid Detection of Shiga Toxin-producing Escherichia coli by Immunochromatography Combined with Asymmetric Polymerase Chain Reaction

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