单核细胞增生李斯特菌PCR鉴定技术的研究

目的建立单核细胞增生李斯特菌PCR鉴定技术.方法选择Hly Inl基因设计扩展片段分别为300～400bp、600～700bp的二对特异性引物,进行PCR扩增.结果单增李斯特菌标准菌株及本实验室分离保存的单增李斯特菌均扩增出两条明显条带,其它食品中常见致病菌及非单核细胞增生李斯特菌均不见扩增条带.52株李斯特菌生化符合菌中,有30株菌同时扩增出两条明显条带,与Mini-VIDSA测定法比较,两者符合率达92.31%.结论本实验建立的PCR鉴定技术为一种简便、快速、敏感、特异的单增李斯特菌鉴定方法.

Study on the Detection of Listeria Monocytogenes with PCR technique

Objective To establish a detection method for Listeria monocytogenes (LM) with PCR technique.Methods The Hly and Inl genes were amplified by designing special primers, and the fragments were 300 400bp and 600 700bp long, respectively.Results These two fragments could be amplified in genomic DNA extracted from standard LM strain or LM strain that our lab isolated and stored, but were not found in other pathogenic bacterium. Among 52 strains conformed to biochemistry reaction of Listeria, 30 strains amplified two fragments simultaneously. As compared to the mini VIDSA detection method, the coincidence rate was 92 31%.Conclusions The PCR technique is a simple, rapid, sensitive and specific method for detecting LM.