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为了解上海地区犬细小病毒的流行及其变异情况,从疑似犬细小病毒感染的死亡犬采集各种组织,无菌处理病料后同步接种F81细胞,盲传至第5代出现细胞病变,从肠组织中分离出1株病毒。采用电镜进行病毒形态学观察,以及HA、HI和PCR等方法鉴定病毒,结果证实为犬细小病毒,命名为CPV-SH15。该病毒的血凝效价为2~9,病毒TCID_(50)为10~(5.2)/m L,测得病毒编码区基因序列的全长为4 869 bp,与上海分离毒株CPV/CN/SH1/2013的同源性为99.8%,亲缘关系较近,说明上海地区的犬细小病毒的遗传变异并不明显。对其VP2基因的氨基酸序列进行分析,确定该细小病毒属于New CPV-2a亚型。该研究为犬细小病毒遗传进化的进一步研究奠定了基础,从而为犬细小病毒病疫苗的研制以及防控提供参考。
In order to understand the prevalence and variation of canine parvovirus in Shanghai, various tissues were collected from dogs suspected of canine parvovirus infection. F81 cells were inoculated synchronously after aseptic treatment of disease materials. One strain of virus was isolated from intestinal tissue. Morphological observation of the virus by electron microscopy, as well as HA, HI and PCR identification of the virus, the results confirmed as canine parvovirus, named CPV-SH15. The titer of the virus was 2 to 9 and the TCID 50 of the virus was 10 to 5.2. The total length of the gene coding for the virus was 4 869 bp, / SH1 / 2013 was 99.8% homologous, which showed that the genetic variation of canine parvovirus in Shanghai was not obvious. The amino acid sequence of VP2 gene was analyzed and the parvovirus belonged to New CPV-2a subtype. This study lays the foundation for the further research on the genetic evolution of canine parvovirus, so as to provide a reference for the development and prevention and control of canine parvovirus vaccine.