单增李斯特氏菌快速鉴定技术的研究

The Study on Listeria Monocytogenes PCR Detection

目的：建立单增李斯特氏菌快速鉴定技术；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：单增李斯特氏菌标准菌株及本实验室分离保存的单增李斯特氏菌均扩增出两条明显条带,其它食品中常见致病茵及非增李斯特氏茵均不见扩增条带．52株李斯特氏菌生化符合菌中,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%(P＞0.5；X2=0.5)．结论：本实验建立的PCR鉴定技术为一种简便、快速、敏感性强、特异性高的单增李斯特氏菌鉴定方法．

[Objective] To establish a method for detecting Listeria Monocytogenes by PCR. [Methods] Amplify Hly, Inl genes by PCR, primer segements in the progress are 300-400bp, 600-700bp,respectively. [Results] The standard Listeria strain and the Listeria strain that isolated and stored in zjcdc lab have amplified two significant bands, however, unListeria strains and pathogenic bacteria from food haven＇ t.30 of 52 bacteria strains that conform to biochemistry reaction of Listeria amplified two significant bands simultaneously.Results of PCR method agreed with those of mini-VIDSA detection by the rate of 92.31（P〉0.5; X2=0.5）. [Conclusions] This assay is a simple, rapid, sensitive and specific method to detect Listeria Monocytogenes.