1株蝙蝠来源细小病毒的分离鉴定

Isolation and identification of a parvovirus from bats

目的对1株蝙蝠来源病毒进行分离鉴定,进行简单的基因分析。方法从云南沧源县8个居民地附近捕获50只棕果蝠并制作组织标本,采用细胞培养分离病毒,采用cDNA-RAPD方法扩增序列,采用分子克隆进一步扩增序列,将序列在NCBI上进行BLAST比对,采用DNAMAN5.0进行同源性分析,采用MEGA 5建序列进化树。结果 50份接毒细胞中,6份细胞出现细胞病变,cDNA-RAPD扩增得到1份阳性片段,测序大小为614 bp,经比对该病毒株是细小病毒科细小病毒亚科博卡病毒属牛细小病毒种的一个毒株。结论成功分离并鉴定该毒株,确定该病毒是牛细小病毒的一个新的基因型。

Objective To isolate and identify a virus from bats and to make simple gene analysis. Methods 50 rousettus leschenaultias were captured from the surrounding areas of 8 residences in Cangyuan County,Yunnan Province and some of their organs were collected;cell culture was adopted to isolate the virus;the sequence of the virus was first amplified by cDNA-RAPD and then further amplified by molecular cloning;the sequence was compared in the NCBI;the homology analysis of the virus was carried out by DNAMAN5. 0 and the phylogenetic analysis was executed by MEGA 5. Results Fifty cells were infected and six cells turned to be CPE and a positive gene fragment was obtained by cDNA-RAPD. The sequencing result indicated that the fragment was 614 bp long. The virus was identified to be one of the Bovine parvovirus of Bocavirus of Parvovirinae of Parvoviridae. Conclusions The isolation and identification of the virus was successful and the virus was confirmed to be a new genotype of Bovine parvovirus.