单核细胞增生性李斯特氏菌荧光定量PCR快速检测方法的建立及初步应用  

Establishment and preliminary application of real-time fluorogenic quantitative PCR assay to rapid detection of Listeria monocytogenes

作　　者：关棣锴[1,2] 胡文忠[2] 朴永哲[2] 陈晨[2] 姜爱丽[2] 王运照 

机构地区：[1]大连工业大学食品学院,辽宁大连116034 [2]大连民族学院生命科学学院,辽宁大连116600

出　　处：《食品工业科技》2014年第20期49-52,56,共5页Science and Technology of Food Industry

基　　金：国家自然科学基金项目(31172009;31340038);国家科技支撑计划项目(2012BAD38B05);大连市科技计划项目(2012E13SF106);大连市金州新区科技计划项目(2012-A1-049)

摘　　要：以单核细胞增生性李斯特氏菌的hly基因为靶基因,用Primer 5.0设计了特异性兼并引物,建立荧光定量PCR检测方法。并用阳性质粒标准品、不同菌株对该方法的特异性、灵敏度进行了验证,且在人工污染鲜切甜瓜上对该方法进行了初步应用。结果表明,建立的检测方法特异性强、灵敏度高,对阳性质粒标准品的灵敏度为7.69×10copies/μL,对人工污染的鲜切甜瓜中单核细胞增生性李斯特氏菌的最低检出限为3.11×10cfu/mL。Hly gene of Listeria monocytogenes was using as the target gene, a pair of degenerate primers were designed with Primer 5.0,and the method of real-time fluorogenic quantitative PCR assay to detection of Listeria monocytogenes was established. The specificity and sensitivity of plasmid standard and different strains were verified,its preliminary application in Artificial inoculation fresh-cut melon was realized. The results showed that the method had good specificity and sensitivity,and sensitivity for plasmid standard was, 7.69x 10copies/μL,the minimum detection limit of artificial inoculation fresh-cut melon was 3.1 lx 10cfu/mL.

关 键 词：单核细胞增生性李斯特氏菌 荧光定量PCR hly 鲜切甜瓜 

分 类 号：TS07[轻工技术与工程]