犬细小病毒SH14株的分离鉴定及其VP2基因的序列分析

Isolation and identification of canine parvovirus SH14 strain and sequence analysis of its VP2 gene

为分离犬细小病毒(canine parvovirus,CPV),用疑似犬CPV感染的病犬肠组织研磨液接种猫肾细胞(CRFK),进行病毒分离,并利用常规病毒学和分子生物学方法以及动物回归试验鉴定分离的病毒,并对其VP2基因序列进行测定及分析。结果显示,该病毒能在CRFK细胞上良好生长并产生细胞变圆、脱落等特征性细胞病变;细胞培养物能扩增出CPV的特异性基因组;对病毒的VP2基因序列和推导的氨基酸序列进行分析,结果表明,本研究的分离毒株属于New CPV-2a型,与CPV SH-2/2011株的亲缘关系最近,其核苷酸同源性和氨基酸同源性分别为99.83%和100%。上述结果表明,成功分离到1株New CPV-2a型犬细小病毒,将其暂命名为SH14株。本研究结果为进一步研究我国犬细小病毒的进化和变异,以及疫苗的研制奠定了物质基础。

In order to isolate a strain of canine parvovirus（CPV）,the intestinal tract slurry of dog suspected infect CPV was inoculated into CRFK cells,and we used methods of virology,molecular biology and animal regression experiment to identify the virus,and sequenced VP2 gene.The results showed that the virus can grow on CRFK cells and produce cytopathic effect,including cell rounding,fall of and other stable cytopathic effect;and cell culture can amplify specific bands,and analyzed VP2 gene,the strain belongs to New CPV-2atype,and the identity of nucleotides with strain of CPV SH-2/2011 was 99.83% and amino acids homology was 100%.In a word,a canine parvovirus was successfully isolated and named as SH14 strain.This study result laid the foundation for studies of the evolution and mutation of CPV and development of vaccine.