基于toxR基因病原哈氏弧菌PCR检测方法的建立  

Development of PCR Method for Detection of Pathogenic Baterium Vibrio harveyi Based on ToxR Gene

作　　者：姚东瑞[1] 朱明[1] 陈丽[1] 毕可然[1] 史爱琴[1] 张晓君[1] 

机构地区：[1]淮海工学院海洋学院,江苏省海洋资源开发研究院,江苏连云港222005

出　　处：《水产科学》2011年第9期559-562,共4页Fisheries Science

基　　金：江苏省水产三项工程项目(PJ2010-58);科技部科技人员服务企业行动项目(2009GJC10044);江苏省自然科学基金资助项目(BK2009163)

摘　　要：基于toxR基因序列设计检测哈氏弧菌的1对特异性引物,通过PCR反应条件优化、PCR反应特异性及敏感性检测,建立一种哈氏弧菌的特异性分子检测方法。试验结果表明,该引物可使哈氏弧菌扩增出大小为390 bp的toxR基因片段,3种水产动物病原弧菌未扩增出任何条带,敏感性检测结果为该PCR反应最低能检测出0.375 ng/μl的哈氏弧菌基因组DNA。该试验建立的基于toxR基因的哈氏弧菌的PCR检测方法可用于哈氏弧菌引起的水产动物疾病的快速诊断及海产品安全检测。In this study,a specific method for detection of pathogenic bacterium Vibrio harveyi,a serious pathogen in aquaculture industry,was established by designing one pair of specific primers based on the toxR gene sequences,by optimizing the PCR reaction condition,and by studying the specificity and sensitivity.The results showed that the PCR primers amplified 390 bp toxR gene fragment from the chromosomal DNA of V.harveyi,and no positive reaction was detected in 3 other pathogenic Vibrio species tested.The PCR reaction sensitivity showed that the primers detected V.harveyi at a level of as few as 0.375 ng/μl.The PCR protocol amplifying toxR gene fragment of the V.harveyi was established for an useful method in the specific and rapid diagnose of the disease caused by V.harveyi.

关 键 词：toxR基因 哈氏弧菌 PCR检测 

分 类 号：S917.1[农业科学—水产科学]