摘要
为研究白斑综合征病毒在湖北省克氏原螯虾(Procambarus clarkii)中的分子流行病学特征,探讨WSSV湖北各毒株间的遗传进化关系,对2016年4月至2016年6月在湖北各克氏原螯虾主要养殖区采集的80份样品用套式PCR方法进行WSSV检测,将阳性样品进行变异区ORF14/15和ORF23/24的基因扩增,连接转化克隆的目的基因,通过测序分析比较湖北各毒株在变异区ORF14/15及ORF23/24的差异性。结果显示:80份样品中有19份检测为阳性,阳性率为23.75%,各地区均存在不同程度的感染,其中有7个地区的毒株在ORF14/15区扩增出2种大小不同的片段,分别为1442bp和630bp,另有4份毒株在ORF23/24区扩增出大小为594bp的片段,与GenBank已公布的标准毒株相比,湖北毒株在ORF14/15和ORF23/24存在大片段缺失。
White spot sydrome virus(WSSV) is the most important reason for lage economic losses in shrimp farming. variable regions have been employed in a number of molecular epidemiology studies of WSSV. Here, we use the major variable loci(ORF14/15 and ORF23/24)as marks of the prevalence and evolution of WSSV. In this study, 80 Procambarus clarkii collected samples from April 2016 to June 2016 in Hubei were detected by nested PCR, 19 sampls tests positive,the infection of the WSSV is 23.75%.These positive samples were genotyped by PCR with specific primers of ORF14/15 and ORF23/24,the amplications were used to clone and sequence. The results of pairwise and multiple alignment analysis showed that there was fragment deletions in the two variable regions. There were 1442 bp amplified fragments in isolates GA1,HM2, and HM3 in ORF14/15 variable region, while 631 bp were amplified in isolates GA2, SZ, XZ,and JM. The ORF23/24 variable region was 606 bp in length in 4 strains and carried a 10970 bp deletion relative to WSSV-TW.
引文