GeXP多重聚合酶链式反应法检测5种常见食源性致病菌  

Detection of 5 kinds of common foodborne pathogens by GeXP multiplex polymerase chain reaction

作　　者：杨梦婕[1] 任佳[2] 李洋[1] 马学军[1] YANG Mengjie;REN Jia;LI Yang;MA Xuejun(National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;The First Affiliated Hospital of Zhengzhou University,Henan Zhengzhou 450052,China)

机构地区：[1]中国疾病预防控制中心病毒病预防控制所,北京102206 [2]郑州大学第一附属医院,河南郑州450052

出　　处：《中国食品卫生杂志》2020年第4期386-390,共5页Chinese Journal of Food Hygiene

基　　金：国家传染病重大专项(2017ZX10104001-002,2018ZX10711001,2018ZX10713-002)。

摘　　要：目的建立一种基于GeXP(GenomeLabTM eXpress Profiling)遗传分析系统的5种常见食源性致病菌检测的新技术。方法针对5种常见食源性致病菌(沙门菌、大肠埃希菌O157∶H7、单核细胞增生李斯特菌、志贺菌和副溶血性弧菌)分别进行基因序列比对(invA、rfbE、PfrA、IpaH、tlh基因),设计特异性引物,建立并优化GeXP多重聚合酶链式反应(PCR)体系,评价其特异性和灵敏性,并初步应用于未知菌株和人工污染样品的检测。结果GeXP多重PCR法可实现在5 h内同时对5种常见食源性致病菌进行检测,且检测灵敏度可低至103 CFU/mL。多重体系中各引物对各目标菌有较强特异性,未检出其他非目标菌。结论本方法可有效提高检测效率,为食源性致病菌的快速高通量检测提供了参考思路。Objective To establish a new and rapid GeXP(GenomeLabTM eXpress Profiling)based multiplex polymerase chain reaction(PCR)assay for the detection of five common foodborne pathogens.Methods Nucleotide sequences of specific gene(invA,rfbE,PfrA,IpaH,tlh)of the five pathogens(Salmonella,Escherichia coli O157∶H7,Listeria monocytogenes,Shigella,Vibrio parahaemolyticus)were obtained and compared.The primers were then designed and the multiplex PCR assay was evaluated.Optimized assay was further validated with the detection of the unknown strains and artificially contaminated samples.Results The GeXP multiplex PCR with five sets of specific primers can be used to detect five foodborne pathogens simultaneously within 5 hours.The specificity was examined by specimens confirmed previously.The detection limit was 103 CFU/mL.Conclusion The results suggested this GeXP multiplex PCR assay was a fast,high throughput test for foodborne bacterial pathogens.

关 键 词：GeXP多重聚合酶链式反应 沙门菌 大肠埃希菌O157∶H7 单核细胞增生李斯特菌 志贺菌 副溶血性弧菌 检测 

分 类 号：R155[医药卫生—营养与食品卫生学]