多种食源性致病菌检测的多重PCR方法的研究  

Study of A Multiplex PCR Method for the Detection of Foodborne Pathogen

作　　者：万志刚[1] 汤慕瑾[1] 吕敬章[1] 罗志军 洪小柳[1] 马淑棉[1] 

机构地区：[1]深圳出入境检验检疫局食品检验检疫技术中心,广东深圳518045 [2]深圳湾出入境检验检疫局,广东深圳518054

出　　处：《现代生物医学进展》2012年第11期2177-2181,共5页Progress in Modern Biomedicine

基　　金：深圳出入境检验检疫局科技项目(SZ2007004)

摘　　要：目的:利用多重PCR技术,建立可以同时检测多种食源性致病菌的多重PCR方法。方法:分别选择沙门氏菌invA基因,志贺氏菌的ipaH基因,单核细胞增生李斯特氏菌的hlyA基因,大肠杆菌O157:H7的eaeA基因,副溶血弧菌的toxR基因,设计多重PCR引物,建立多重PCR检测体系,并对该体系进行特异性和灵敏度实验。结果:通过对19株菌株进行实验,所有的目标菌株均为阳性,而其余菌株为阴性。对多重PCR体系的灵敏度进行考察,沙门菌的灵敏度为5000 CFU/mL;志贺氏菌的灵敏度为5500CFU/mL;单核细胞增生李斯特氏菌的灵敏度为5200 CFU/mL;O157:H7的灵敏度为5000CFU/mL;副溶血弧菌的灵敏度为6300CFU/mL。结论:建立的多重PCR体系能实现多种致病菌同时检测。Objective： To develop a multiplex PCR method to detect five food borne pathogenic microorganisms simultaneously.Methods： Primers specific for invA gene of Salmonella spp.,ipaH gene of Shigella spp.,hlyA gene of Listeria monocytogenes,eaeA gene of Escherichia coli O157：H7 and toxR gene of Vibrio parahaemolyticus were designed,and the specificity and sensitivity of the devel-oped method was further verified.Results： A collection of 19 strains was examined,all target strains were detected.In contrast,none of the non-target strains yielded the specific amplification product.The sensitivity of the multiplex PCR system was 5000 CFU/mL for Salmonella cultures,5500 CFU/mL for Shigella cultures,5200 CFU/mL for Listeria monocytogenes cultures,5000 CFU/mL for Es-cherichia coli O157：H7 cultures and 6300 CFU/mL for Vibrio parahaemolyticus cultures.Conclusions： The multiplex PCR method in present study can be applied in practice.

关 键 词：多重PCR 食源性致病菌 

分 类 号：R378[医药卫生—病原生物学]