淡水小龙虾弧菌分离株毒力基因的筛选与检测方法的建立

A Method for Screening and Detection of Virulence Genes of Vibrio Isolates from Freshwater Crayfish

目的:筛选湖北淡水小龙虾弧菌分离株毒力基因并建立一种特异、灵敏的检测方法。方法:应用分子生物学——聚合酶链式反应(polymerase chain reaction,PCR)技术,根据文献记载的弧菌属中6种不同的分离株CTX基因元件中的ctxA、RS1、zot基因,VPI毒力岛中的tcpA、toxT基因,RTX基因簇中的rtxC,以及toxR、T139、tcpH、toxRS、toxS、ctxAB基因序列设计PCR引物建立PCR检测方法。结果:该方法能够有效地对弧菌分离株毒力基因进行检测,简便快速、特异性强、检测周期短。

A specific and sensitive PCR method for the detection of Vibrio was established.The ctxA,zot and RS1 genes of CTX genetic element,tcpA and toxT genes of VPI pathogenic island,rtxC gene of RTX cluster,and toxR,T139,tcpH,toxRS,toxS and ctxAB genes in 6 Vibrio isolates reported in the literature were used to design specific primers for PCR.The results indicated that the established method was simple,easy,specific and time-saving.As a result,it can effectively and efficiently detect virulence genes of Vibrio strains.