猪细小病毒HLJ-DN株分离鉴定

Isolation and characterization of porcine parvovirous strain HLJ-DN

按照常规病毒分离方法,从黑龙江省某猪场怀孕母猪所产死胎的肝脏中分离到一株病毒。利用ST细胞进行培养,通过血细胞凝集试验、间接免疫荧光试验及电镜观察等对新分离毒株进行鉴定。结果表明,该病毒能在ST细胞上生长并引起明显拉网状细胞病变,按Reed-Muench法计算出TCID50为10-4.6·0.1 mL-1;通过电镜观察可见直径约为20 nm无囊膜的病毒粒子,该病毒能凝集0.5%的豚鼠红细胞,免疫荧光试验显示在细胞上出现明显荧光,能与特异性抗体发生反应。对VP2基因进行扩增及测序并与GenBank已发表PPV毒株序列进行比对分析,结果显示该毒株与WB-143株在同一个分支里,同源性高达到99.8%,说明分离的病毒为猪细小病毒,命名为HLJ-DN,该病毒的分离为研究当地猪细小病毒的流行、预防和治疗奠定。

This research separates a virus from the liver of natural infection pregnant sows stil birth from a farm in Heilongjiang Province, this virus can grow in the ST cells and lead to an obvious pathological changes in reticuLar cell. According to the Reed Muench method we can calculated the TCID50 is 10-4.6·0.1 mL-1; By electron microscope we can observe some virus particles without posterior capsuLe and their diameter is about 20 nm, the virus can agglutinate 0.5% RBC of guinea pig, immunofluorescence test showed a significant fluorescence in the cells, they can react with specific antibody, at the same time we do the VP2 gene amplification and sequencing, we find the sequence alignment is correct. The results show the strain and WB-143 strain are the same branch, up to 99.8% homology. These results il ustrated that the separation virus is porcine parvovirus, it named HLJ-DN.