水产品中致病性弧菌PCR快速检测研究  

Study on Rapid Detection Technique of Polymerase Chain Reaction on Pathogenic Vibrio spp.in Aquatic Products

作　　者：冯家望[1] 王小玉[1] 李丹琳[1] 唐食明[1] 刘锐[1] 

机构地区：[1]珠海出入境检验检疫局,珠海519015

出　　处：《中国国境卫生检疫杂志》2007年第6期383-386,共4页Chinese Journal of Frontier Health and Quarantine

基　　金：珠海出入境检验检疫局科研基金项目:(ZH2003-7)

摘　　要：〔目的〕建立一种快速、准确、特异的方法,快速检测和鉴定出食品中的非致病性和致病性霍乱弧菌与副溶血性弧菌。〔方法〕针对霍乱弧菌的种特异性基因ompW、毒力基因tcpA、ctxA和副溶血性弧菌的种特异性基因tl、毒力基因tdh设计引物,建立PCR检测体系。〔结果〕检测结果显示,该方法能够特异性检出副溶血性弧菌或霍乱弧菌,并进一步确定其是否携带tdht、cpA或ctxA毒力基因,检测的灵敏度可达到5×101cfu/ml菌浓度。〔结论〕与传统方法比较,该方法快速、特异、灵敏,能在较短时间内对大量水产品样品进行检测。Objective To establish a polymerase chain reaction method to detect Vibrio cholera and Vibrio parahaemolticus in aquatit products. Method The virulence genes of tcpA, ctxA and tdh were selected as target sequences, ompW gene was selected as a specific genomic marker for Vibrio cholera and tl for Vibrio parahaemolyticus. The primers were designed and synthesized for rapid detection and identification of pathogenic Vibrios and other closely related Vibrio. Result It was shown that this method can specifically identify Vibrio cholera and Vibrio parahaemolyticta~ and confirm whether they contain tdh, tcpA or ctxA virulence gene or not. The sensitivity of the method was 5×10^3 cfu/ml. Conclusion In contrast to the conventional culture technique, the RCR method is more rapid, specific and sensitive. It is able to detect pathogenic Vibrios from a lot of food in a shorter period and is well suited for entry-exit inspection of aquatic products.

关 键 词：聚合酶链式反应 致病性霍乱 弧菌 副溶血性 毒力基因 水产品 

分 类 号：R155.5[医药卫生—营养与食品卫生学]