| 摘要: |
| 采集北京地区疑似细小病毒(CPV)感染病犬血样粪便,采用同步培养法接种猫肾细胞(CRFK)分离病毒,通过PCR试验、HA试验、HI试验、IFA试验和电镜观察鉴定为犬细小病毒,命名为CPV-DD株。分离毒株可扩增出特异DNA片段(1163bp)；盲传至第2代时,病毒液可凝集猪红细胞,血凝价为1:8,其血凝性能被特异性抗体抑制；IFA可见特异性亮绿色荧光,TCID50为103.0／0.1mL；电镜观察可见20nm左右的病毒颗粒,VP2基因序列分析显示该毒株为CPV-2a型,氨基酸序列在324位出现Try→Ile。 |
| 关键词: 犬细小病毒 VP2 分离 鉴定 |
| DOI： |
| 投稿时间：2011-07-18修订日期：2011-08-25 |
| 基金项目: |
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| Isolation and Identification of Canine Parvovirus DD Strain and Sequence Analysis of the VP2 GeneLi Run, Ling Ning, Yin Chunsheng, Xue Qinghong, Zhi Haibing |
| lirun |
| (China Institute of Veterinary Drug Control) |
| Abstract: |
| A strain of canine parvovirus(CPV)was isolated from bloody feces of an ill puppy suspected of parvovirus infections in Beijing. The strain, designated as CPV-DD, was isolated by inoculating CRFK cells in synchronism and was identified as serotype CPV-2a by PCR, the HA, IFA,electron microscopy observation and sequencing of VP2 gene. A unique Ile-324 in VP2 of DD strain was found, contrasting to a Try-324 in the VP2 of other strains. After 2 passages in CRFK cells, the infected cell cultures could agglutinate red blood cells of pigs at the titer of 1:8. The haemagglutination could be inhibited by monoclonal antibody specific to CPV. Specific fluorescent in the nucleus of the infected cells was observed by IFA assay and the TCID50 of virus fluid was titred as 103.0/0.1ml. Concentrated virus particles with a diameter of 20nm were observed by transmission electron microscope. |
| Key words: canine parvovirus VP2 isolation identification |
