基于PCR-免疫试纸条法的空肠弯曲菌快速检测

Rapid detection of Campylobacter jejuni based on PCR-immune strip method

李鑫岗 ¹刘志勇 ²申进玲 ³郭德华 ³赵丽娜 ³于婕 ³陈瑜 ³周甜甜 ¹王璇 ¹徐博捷 ¹只帅¹

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作者信息

1. 宁波大学医学部公共卫生学院,宁波 315211
2. 上海市徐汇区疾病预防控制中心,上海 200237
3. 上海海关动植物与食品检验检疫技术中心,上海 201210
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摘要

空肠弯曲菌引起的食源性腹泻疾病已经成为突出的公共卫生问题之一,快速检测食品中空肠弯曲菌污染对于保障人类健康具有重要意义.本研究以空肠弯曲菌lpxA基因序列为检测靶基因,通过聚合酶链式反应(polymerase chain reaction,PCR)扩增产物上标记的地高辛和异硫氰酸盐(isothiocyanate,ITC)分子与试纸条上抗体特异性结合,建立了一种PCR-试纸条检测方法.所有空肠弯曲菌标准菌株均为阳性结果,其他弯曲菌以及食源性病原菌均为阴性结果,检测特异性为100%;最低检出限为3.1×103 CFU/mL;在对食品的实际检测中,该方法与现有国标检测方法结果一致.将本方法用于空肠弯曲菌的检测降低了成本,简化了操作步骤,为食源性空肠弯曲菌检测提供了一种实用、简便的手段.

Abstract

Food-borne diarrheal disease caused by Campylobacter jejuni(C.jejuni)has become one of the prominent public health problems.Rapid detection of C.jejuni contamination in food is of great significance for protecting human health.In this study,the lpxA gene sequence of C.jejuni was used as the target gene,a PCR-test strip detection method was established by binding digoxin and isothiocyanate(ITC)molecules labeled on polymerase chain reaction(PCR)products to antibodies on the strip.All standard strains of C.jejuni were positive by our method,while other Campylobacter species and food-borne pathogens were negative result.The detection specificity was 100%,and the detection limit was 3.1×103 CFU/mL.This method is consistent with the results of the current national standard detection methods.The application of this method for the detection of C.jejuni would reduce the cost,simplify the operation procedure,and provide a practical and easy method for the detection of food-borne C.jejuni.

关键词

空肠弯曲菌/聚合酶链式反应/免疫试纸条

Key words

Campylobacter jejuni/polymerase chain reaction/immune test strip

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基金项目

上海市技术标准专项项目(23DZ2203700)

宁波市自然科学基金(2023J074)

宁波市公益性研究计划项目(2023S071)

出版年

2023

生命的化学

中国生物化学与分子生物学会

生命的化学

CSTPCD

影响因子：0.404

ISSN：1000-1336

参考文献量2

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