虎源副流感病毒5型的分离鉴定及全基因序列分析

Isolation,identification and complete gene sequence analysis of tiger parainfluenza virus type 5

本研究旨在了解虎源副流感病毒5型(parainfluenza virus 5,PIV5)的遗传变异情况,阐明其基因组与遗传进化等特征。从广东省某动物园疑似虎PIV5感染的病料处理后接种Vero细胞并通过一系列的鉴定,将分离的毒株命名为HMZ。根据GenBank中PIV5基因序列,设计包含全长的12对特异性引物,并扩增其全基因组。将扩增产物克隆到pMD-18T载体上,对鉴定为阳性的重组质粒送检测序,并对基因全序列进行同源性分析及遗传进化分析。结果显示,HMZ与各病毒株全基因序列同源性为97.2%～99.8%。其中与1168-1 (KC237064.1)、ZJQ-221 (KX100034.1)以及Mammalian rubulavirus 5 (KY685075.1)亲缘性最高,为99.8%;而与D277的亲缘性最低,为97.2%。全基因系统进化树分析结果表明,HMZ与ZJQ-221、Mammalian rubulavirus 5属于同一分支。

The purpose of this study is to understand the genetic variation of parainfluenza virus5(PIV5)and to elucidate its of genome and genetic evolution charactertion.Vero cells were inoculated with sample suspect infected PIV5 of a tiger from a zoo in Guangdong Province.The isolated strain was named as HMZ after a series of identification.According to the sequence of PIV5 gene in Gen Bank,Twelve overlapping fragments covering the whole viral genome were designed and the full-genome were amplified.The amplified products were cloned into p MD-18 T vector,then sequenced the positive recombinant plasmids.The whole gene sequence was analyzed using homology analysis and genetic evolution analysis.The results showed that HMZ was homologous with 97.2%-99.8%of all virus strains.Alignment analysis indicated that HMZ had the most proximate consensus(99.8%)to 1168-1(KC237064.1),ZJQ-221(KX100034.1)and Mammalian rubulavirus 5(KY685075.1),while the least consensus(97.2%)to D277(KC237065.1).Phylogenetic tree analysis revealed that HMZ strain,ZJQ-221 strain and Mammalian rubulavirus 5 belonged to the same branch.