犬细小病毒山东流行株分离鉴定及其全基因组测序分析

Isolation,identification and geonomic analysis of canine parvovirus in Shandong Province

为了解山东地区犬细小病毒的流行及其变异情况,应用F81细胞从山东地区送检的发病犬粪便中分离出4株细小病毒,根据PCR和电镜技术对其进行鉴定,并对分离株的全基因组进行克隆测序与序列分析。结果表明：所分离到的4株病毒均能在F81细胞上产生明显的细胞病变,经PCR及电镜观察鉴定为犬细小病毒,分别命名为QN1/QN2/QN3/QN4株。全基因组分析结果显示,除QN3株的基因组全长为4 756 nt外,其余3株均为4 757 nt;4个分离株之间全序列核苷酸同源性为99.31%,与GenBank登录的10株具有代表性的CPV核苷酸序列比对,同源性为98.2%-99.9%;VP2基因的核苷酸序列同源性为98.5%-99.9%,氨基酸序列同源性为98.1%-100%;表明各分离毒株的亲缘关系较近,同属于NewCPV-2a亚型。

To determine the epidemiology of canine parvovirus in Shandong,four strains of parvovirus were isolated using F81 cells from feces collected from sick dogs,the CPV strains were confirmed by PCR and transmission electron microscope and the full genome nucleotide sequence was cloned, sequenced,and analyzed. The results show that four isolates produced specific CPE in F81 cells, further identified by PCR and electron microscope,named as QN1/QN2/QN3/QN4,respectively. The full genome sequence analysis results indicated that the genome was composed of 4 757 nucleotides,with exception of one isolate（QN3） showing 4 756 nt.Homology of the nucleotides sequence is 99.31% among this 4 isolates. Comparative analysis reveals that the isolates have high homology with the genomic sequence of 10 representative strains, ranging from 98.2%~99.9%.The VP2 gene sequence and the deduced amino acid sequence are 98.5%~99.9% and 98.1%~100%, respectively. It Shows this isolates have close genetic relationship,belonging to the same New CPV-2a subtype.