犬细小病毒上海分离株全基因的克隆及进化树分析

COMPLETE GENOMIC SEQUENCE ANALYSIS OF A SHANGHAI LSOLATE OF CANINE PARVOVIRUS

为研究上海地区犬感染细小病毒(Canine parvovirus,CPV)的基因特点和基因型,参考GenBank上细小病毒3个基因型的全基因设计6对扩增引物进行PCR,分段扩增出目标片段,克隆于T载体上,测定序列并拼接,用MEGA软件、NJ法构建进化树。结果显示,犬细小病毒全基因为4 758bp,命名为CPV-SH,提交到GenBank上的序列号为FJ792845,与GenBank中4种基因型犬细小病毒的VP2基因核苷酸序列同源性比较发现,CPV-SH克隆与2a亚型犬细小病毒核苷酸同源性为最高99.7%,与2型、2b亚型、2c亚型的同源性分别为98.8%、99.5%、99.5%。CPV-SH属于2a亚型犬细小病毒,与国内北京分离株CPV/BJ082亲缘关系最近,同源性为99.9%。

To study the genomic organization and genotype of a Shanghai isolate of Canine parvovirus （CPV-SH）, primers were designed on the basis of the complete genomic sequence of 3 Canine parvovirus serotypes in the GenBank, and the genomic sequence was amplified from the isolate by PCR. The PCR products were cloned into T vector and sequenced. Phylogenetic tree was constructed by the Neighborjoining method with Mega 4 software. The complete genomic sequence of CPV-SH was 4 758 bp in length. The full genome sequence was deposited in GenBank under access no. FJ792845. Comparison of this sequence with that of VP2 gene of 3 Canine parvovirus serotypes in GenBank revealed high homology with genotypes 2a,2, 2b and 2c （99.7G, 98. 8G, 99. 5%, and 99. 5%）. Genetic analysis of 20 other Chinese isolates also found that CPV-SH belonged to genotype 2a and shared 99. 9% homology with a Beijing isolate （CPV/BJ082）.