五株猪流行性腹泻病毒的分离鉴定及序列分析  

Isolation and Identification of Five Porcine Epidemic Diarrhea Virus Strains and Sequence Analysis

作　　者：王贵华[1,2] 于萍萍[1,2] 满坤[1,2] 吴珊珊[1,2] 刘明明[1,2] 孟萌[1,2] 赵亚荣[1,2] 

机构地区：[1]北京大北农科技集团股份有限公司动物医学研究中心,北京100195 [2]北京市畜禽生物制品工程技术研究中心,北京100195

出　　处：《中国兽药杂志》2016年第7期13-19,共7页Chinese Journal of Veterinary Drug

摘　　要：为研究猪流行性腹泻病毒的流行情况并分析该病毒的培养特性,对甘肃、北京、河北、山西、浙江五个猪场送检的疑似猪流行性腹泻病猪小肠内容物,用RT-PCR方法进行检测,证明为猪流行性腹泻病毒核酸阳性。然后将阳性病料进行传代培养、病毒含量测定以及特异性检验,证明获得五株猪流行性腹泻病毒,分别命名为Ningxia9、Beijing5、Gaocheng8、Shanxi1、Zhejiang08。对分离的毒株进行S基因全长测序及氨基酸序列分析。结果表明：五株分离毒株在盲传19代开始出现明显的细胞病变,盲传到24代,出现稳定的细胞病变且五株分离毒株的24代病毒含量在104.5-5.0TCID50/m L之间;用S蛋白进行进化树分析表明,五株分离毒株与近年来国内流行毒株亲缘关系很近,在同一个进化分支上;分离的五株野毒株之间的氨基酸同源性高达99.2%-99.9%,而这五株野毒与Gen Bank上提交的2011年、2012年分离的野毒株的之间的氨基酸同源性为98.4%-99.9%;实验室分离毒株及国内近年流行毒株均与国内外经典毒株存在一定的共性,即特定位置氨基酸的插入、缺失或置换。In order to research epidemiology and culture specificity of porcine epidemic diarrhea virus（ PEDV）,five feces samples from piglets suffering severe diarrhea in farms from Gansu,Beijing,Hebei,Shanxi and Zhejiang,were confirmed as PEDV positive by RT-PCR. Then these five samples were cultured by vero cell line to 24 passages,the five cultures were titrated and sequenced full long of S gene. Results showed that cytopathic effect（ CPE） appeared from 19 thto 24thpassages,and virus titer were between 104. 5 - 5. 0TCID50/ m L. Sequence analysis showed that the amino acid homology was 99. 2% - 99. 2% among five wild strains,98. 4% - 99. 9%among these five strains and the wild strains on Gen Bank from 2011 to 2012. Furthermore,the isolates,domestic pandemic strains and classic strains had certain commonality such as specific amino acid insertions and deletions or replacement.

关 键 词：猪流行性腹泻病毒 分离鉴定 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]