2种弧菌的实时荧光PCR快速检测  

Study on Rapid Detection of 2 kinds of Vibrio spp.in Food by Real-Time Fluorescence PCR

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

机构地区：[1]珠海出入境检验检疫局,广东珠海519015

出　　处：《中国食品卫生杂志》2007年第6期514-518,共5页Chinese Journal of Food Hygiene

基　　金：珠海出入境检验检疫局科研项目(ZH2003-7)

摘　　要：目的为快速、特异、灵敏的检测致病性弧菌,建立致病性弧菌的实时荧光PCR方法。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设计引物和Taqman荧光探针,建立实时荧光PCR检测方法。结果该方法能够特异性地检出副溶血性弧菌或霍乱弧菌,并进一步确定其是否携带tdh、tcpA或ctxA毒力基因,检测的灵敏度可达到10CFU/ml或0.1716μg/ml(pg/μl)DNA模板浓度。结论该方法特异性强、灵敏度高,适用于食品中致病性弧菌的快速检验。Objective To develop a real-time fluorescence PCR assay to detect Vibrio cholera and Vibrio parahaemolticus in food. 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 primer and Taqman probes were designed and synthesized for rapid detection and identification of pathogenic Vibrios and other closely related Vibrio. Results It is shown that the method could specifically identify Vibrio cholera and Vibrio parahaemolyticus and confirm whether they contain tdh, tcpA or ctxA virulence gene or not. The sensitivity of the assay was 10 CFU/ml or 0.171 6 μg/ml. Conclusion The real-time fluorescence PCR assay could have high sensitivity and specialty, and it could be well suited for detect Vibrio cholera and Vibrio parahaemolticus in food.

关 键 词：聚合酶链反应 弧菌属 弧菌 霍乱 弧菌 副溶血性 毒力 基因 食品 

分 类 号：R117[医药卫生—公共卫生与预防医学]