摘要
为获得1株猪伪狂犬病病毒贵州分离株,将贵阳市某猪场送检的流产胎儿病料经一系列处理后接种Vero细胞,并对分离毒株分别进行了病毒理化特性、电镜形态观察、中和试验、动物接种试验、核酸检测等鉴定。当盲传至第3代时,出现细胞变圆变亮、脱落、呈拉网状特征的细胞病变;分离毒不耐酸和热,对5-碘脱氧尿核苷和有机溶剂氯仿敏感,能被伪狂犬病病毒标准阳性血清中和;透射电镜下可见位于囊泡中呈近圆形、有囊膜、150～180 nm大小的成熟病毒粒子,在胞核中可见衣壳呈晶格状排列的病毒包涵体以及空心、实心2种无囊膜的病毒粒子;将分离毒株接种家兔,能引起家兔奇痒、麻痹死亡;PCR扩增产物经测序证明系PRV gD基因的部分序列。结果表明,成功分离获得了1株猪伪狂犬病病毒贵州株。
To obtain a strain of pseudorabies virus（PRV）,The samples of the porcine aborted fetus isolated from a hoggery in Guiyang were preprocessed,and then,were inoculated into the Veto cells. The inoculated Veto cells showed the typical cytopathogenic effect when cultured to the third generation. The virus was identified by the physicochemical experiment, transmission electron microscopy, neutralization tests, animal inoculation, and viral nucleic acid detection. The results indicated that the virus was sensitive to acid,heat,IUDR and chloroform, and could be neutralized by the PRV positive serum. By transmission electron microscopy,the mature virions with envelope in uesicae were round or ellipse with a diameter of 150 to 180 nm,and the ion in nucleus were hollow or stuffed without envelope,and the capsids were lattice arrangement in mature viral inclusion bodies. The rabbits injected with cell cultures showed the typical clinical syndrome of pruritus,palsy and death in the end. Sequencing results of PCR product indicated that the obtained sequence was gD partial sequence of PRV. All of the results indicated that the PRV strain was isolated successfully. It was named Guizhou strain.
出处
《中国兽医科学》
CAS
CSCD
北大核心
2011年第5期459-463,共5页
Chinese Veterinary Science
基金
贵州省科学技术基金项目(黔科合J字[2007]2065)
贵州省农业科技攻关项目(黔科合NY字[2010]3042)
关键词
猪伪狂犬病病毒
分离
鉴定
porcine pseudorabies virus （PRV）
isolation
identification
