三种常见食源性致病菌的多重PCR快速检测

摘要

In order to establish a rapid, sensitive and specific PCR diagnostic method to detect the three foodbome pathogens including Esche-richia coli,Salmonella typhimurium and Listeria monocytogenes, their specific primers were designed. The specificity and sensitivity of primers were verified by single, duplex and triplex-PCR reaction, and by detecting the foods artificially infected with the three pathogens. The results showed that three foodbome pathogens were amplified purpose fragments, and no other non-specific fragments were amplified. PCR test for the foods artificially infected with the pathogens also received specific segments of the purpose bacteria, and the results were stable. The multiplex PCR method in the present study was highly specific and sensitive. It would provide a fast, simple, and reliable method for the rapid detection of foodbome pathogens, which had significant theoretical and practical meaning.%为建立对3种食源性致病菌—大肠杆菌(Escherichia coli)、沙门氏菌(Salmonella typhimurium)和单增李斯特菌(Listeria monocytogenes)的快速、敏感、特异的多重PCR诊断检验方法,本研究设计了上述3种菌的特异性PCR引物,通过单一、双重及三重PCR反应检测,并对人工感染的饮料和面包2种食品进行验证.结果表明,3种食源性致病菌的单一、双重和三重PCR均成功扩增出目的片断,没有非特异性扩增.人工感染食品的PCR检测也获得了目的菌的特异性片段,并且结果稳定.本研究所建立的多重PCR反应系统具有较好的特异性和灵敏性,为食品中常见致病菌的检测提供了一种快速、简便、可靠的方法,具有重要的理论意义及实践意义.