犬细小病毒HZ0761株的分离与鉴定

采集疑似细小病毒(CPV)感染犬的粪便,采用同步培养法接种胎猫肾细胞(F81)进行病毒分离鉴定。通过PCR检测、HA试验、IFA鉴定、电镜观察和空斑纯化,获得1株犬细小病毒,并命名为HZ0761。感染的F81细胞48h后出现明显的细胞病变;在病料和感染的F81细胞中均扩增出CPV VP2基因的特异性片段(221 bp);病毒液可凝集猪红细胞,血凝价为1∶28,其血凝性能被特异性抗体抑制;IFA可见特异性亮绿色荧光;电镜观察感染的F81细胞核内可见20 nm左右的病毒颗粒;病毒液的TCID50为10-4.8/mL,VP2基因序列分析显示该毒株为CPV-2 a型。

Isolation and identification of canine parvovirus strain HZ0761

To isolate canine parvovirus(CPV),embryonic feline kidney cell line(F81) was inoculated with fecal specimens from dogs suspected of parvovirus infections.One CPV isolate,designated as HZ0761,was purified by plaque formation and characterized by HA test,immunofluorescence assay on F81 cells and electron microscopic observation as well as sequence analysis.The 221 bp product specific for CPV VP2 gene was amplified by PCR from fecal specimen and infected F81 cells in which cytopathic effect(CPE) was found after 48 hours post infection.Visible signals were observed in infected cells by IFA and the TCID50 of virus fluid was titred as 104.8/mL.The viral stock could agglutinate red blood cells of pigs at the titer of 28,and the haemagglutination could be inhibited by monoclonal antibody specific to CPV.The virus particles about 20 nm in size were observed under electron microscope.Moreover,sequencing of VP2 gene indicated that the isolate was CPV-2a type.