级联信号转导系统结合免疫层析法检测大肠埃希氏菌O157:H7

doi:10.7506/spkx1002-6630-20200615-196

食品科学 ›› 2021, Vol. 42 ›› Issue (18): 314-320.doi: 10.7506/spkx1002-6630-20200615-196

级联信号转导系统结合免疫层析法检测大肠埃希氏菌O157:H7

山珊，黄艳梅，赵雪龙，陈文瑶，刘成伟，龙中儿，刘道峰

（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.南昌大学食品科学与技术国家重点实验室，江西南昌 330047；4.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）

发布日期:2021-09-29
基金资助:
江西省教育厅科学技术研究项目（GJJ191708）；江西省卫健委科技计划项目（20201120）；江西省自然科学基金项目（20202BAB206066）；江西省重点研发计划项目（20192BBGL70053）

Detection of Escherichia coli O157:H7 by Cascade Signal Transduction System Combined with Immunochromatography

SHAN Shan, HUANG Yanmei, ZHAO Xuelong, CHEN Wenyao, LIU Chengwei, LONG Zhong’er, LIU Daofeng

(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. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China; 4. Jiangxi Province Key Laboratory of Diagnosing and Tracing of Foodborne Disease, Jiangxi Provincial Center for Disease Control and Prevention, Nanchang 330029, China)

Published:2021-09-29

摘要/Abstract

摘要： 首先通过免疫磁技术实现目标菌的有效富集，然后经同时标记了大量抗体与β-内酰胺酶的胶体金纳米探针介导，利用β-内酰胺酶高效水解青霉素实现检测信号转导及放大，再通过免疫层析法对青霉素含量变化的灵敏甄别，最终实现大肠埃希氏菌O157:H7的超灵敏检测。建立的检测方法对大肠埃希氏菌O157:H7的检测灵敏度为2×102 CFU/mL，相比传统的免疫层析法检测灵敏度提高了570 倍。实现超灵敏检测食源性致病菌的同时规避了双抗夹心免疫层析法中Hook效应的影响，为免疫层析高灵敏准确检测大分子目标物提供了新的思路。

关键词: 信号转导；免疫层析技术；超灵敏检测；大肠埃希氏菌O157:H7

Abstract: For ultrasensitive detection of Escherichia coli O157:H7, the target bacteria were enriched effectively by immunomagnetic separation. Then, colloidal gold nanoprobes simultaneously with a large amount of antibodies and β-lactamase were used as the mediators. β-Lactamase was employed to hydrolyze penicillin efficiently for the transduction and amplification of detection signals. Immunochromatography was applied for detecting the variation in penicillin concentration. The sensitivity of the developed method was 2 × 102 CFU/mL, which was 570 folds higher than that of traditional immunochromatography. In addition, this method avoided the Hook effect in sandwich immunochromatography, which provides a new tool for immunochromatographic detection of macromolecular analytes with high sensitivity and accuracy.

Key words: signal transduction; immunochromatography; ultrasensitive detection; Escherichia coli O157:H7

中图分类号:

山珊，黄艳梅，赵雪龙，陈文瑶，刘成伟，龙中儿，刘道峰. 级联信号转导系统结合免疫层析法检测大肠埃希氏菌O157:H7[J]. 食品科学, 2021, 42(18): 314-320.

SHAN Shan, HUANG Yanmei, ZHAO Xuelong, CHEN Wenyao, LIU Chengwei, LONG Zhong’er, LIU Daofeng,. Detection of Escherichia coli O157:H7 by Cascade Signal Transduction System Combined with Immunochromatography[J]. FOOD SCIENCE, 2021, 42(18): 314-320.

级联信号转导系统结合免疫层析法检测大肠埃希氏菌O157:H7

Detection of Escherichia coli O157:H7 by Cascade Signal Transduction System Combined with Immunochromatography

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