中文摘要：

根据副溶血弧菌种特异性基因（tox R）、耐热直接溶血素基因（tdh）和相对耐热直接溶血素基因（trh）为靶基因分别设计引物,进行PCR扩增及反应条件的优化,建立了检测含有溶血素毒力基因的致病性副溶血弧菌的多重PCR方法。3对引物能分别特异性地扩增出368、269、486 bp的目的片段。副溶血弧菌均能扩增出tox R基因,不含溶血素基因的菌株（tdh-/trh-）仅扩增出1条目的片段,而含不同溶血素基因的致病性副溶血弧菌则分别扩增出2条（tdh＋/trh-或tdh-/trh＋）和3条（tdh＋/trh＋）特异性条带。检测其他非副溶血弧菌的供试菌,则不出现任何扩增条带。人工模拟样品检测结果显示对致病性副溶血弧菌的最低检测浓度为103CFU/m L。结果表明该多重PCR检测方法具有较好的特异性和灵敏性,对检测致病性副溶血弧菌有重要意义。

英文摘要：

In order to identify Vibrio parahaemolyticus that produce toxin genes,a rapid,sensitive and specific multiplex PCR assay was developed in this study.Three pairs of primers were designed to detect species-specific gene（toxR）,heat direct hemolysin gene（tdh） and heat-related hemolysin gene（trh） of V.parahaemolyticus,respectively.The reaction conditions for the multiplex PCR were optimized to establish a rapid,sensitive and convenient method for detection of pathogenic V.parahaemolyticus in aquatic animals.The results showed that the multiplex PCR produced specific amplicons with expected sizes 368 bp,269 bp and 486 bp.The three pairs of primers amplified different V.parahaemolyticus strainss：one band for tdh～-ltrh～- V.parahaemolyticus,two bands for tdh～＋ltrh～- or tdh～-/trh～＋V.parahaemolyticus and three bands for tdh～＋/trh～＋ V.parahaemolyticus.The multiplex PCR did not detect any other bacteria.The detection limit of the multiplex PCR was 1×10～3 CFU/mL for pathogenic V.parahaemolyticus.Development of the multiplex PCR offers an efficient way for high sensitive and special detection of pathogenic V.parahaemolyticus.