3种主要水产病原菌多重PCR检测方法的建立

Multiplex PCR for the detection of three main aquatic pathogens

中文关键词: 鳗弧菌  嗜水气单胞菌  迟缓爱德华氏菌  多重PCR

中文摘要:

      根据目前水产上常见病原菌鳗弧菌Vibrio anguillarum、嗜水气单胞菌Aeromonas hydrophila、迟缓爱德华氏菌Edwardsiella tarda的毒力相关基因，选择具有特异性的鳗弧菌调控毒力蛋白表达的toxR(Positive transcriptional regulator)基因、嗜水气单胞菌中重要的编码气溶素(Aerolysin)基因aerA、迟缓爱德华氏菌中编码分泌系统装置蛋白的基因evpA，分别设计1对特异性引物，建立可同时特异性地检测3种菌的多重PCR检测体系。对反应条件进行优化并测试了其特异性和灵敏度。实际样品检测证明，此多重PCR体系具有良好的可靠性。

英文摘要:

      A multiplex polymerase chain reaction (PCR) was developed for synchronous detection of three major pathogens of aquatic animals, including Vibrio anguillarum, Aeromonas hydrophila and Edwardsiella tarda. Three important and special virulent genes were selected for the multiplex PCR, such as positive transcriptional regulator toxR gene of V. anguillarum, aerolysin gene aerA, which encodes important virulent factor of A. hydrophila, and evpA gene, which encodes the apparatus protein of secretion system T6SS of E. tarda. The reaction condition was optimized and the specificity and sensitivity was tested for this method. The detection for real samples also proved its reliability.