猪圆环病毒河南株的分离鉴定

Isolation and Identification of Porcine Circovirus Henan Strain

【研究目的】为了研究猪圆环病毒的分子生物学特征及其发病机理和致病机制,【研究方法】将疑似猪圆环病毒2型的猪脾脏、淋巴结用胰蛋白酶消化处理后接种PK-15细胞,盲传6代后,病毒已经适应PK-15细胞,【研究结果】然后设计一对特异性引物利用PCR技术扩增出ORF2片段中540bp的基因。第6代细胞病毒液经间接免疫荧光检测表明,在荧光显微镜下可见PCV特征的荧光斑,表明毒株具有感染性。【结论】将病毒纯化后进行电镜观察,病毒粒子直径约17nm,圆形,无囊膜,研究结果表明分离的病毒为猪圆环病毒。

The objective of the prenscnt study was to reserch molecular biology and nosogenesis of porcine eireovirus, isolate porcine eircovirus from pigs and then identify these strains by specific PCR assay. In this experiment spleen and lymph of swine were treated with tupsinase, then inoculated on pk-15 cells line and adapted toeell lines.540 bp gene of OtiF2 segment was applified by PCR technology from pk-15 cell. characteristic PCV fluorescence was discovered in fluorescence microscope by IFA （ indrect fluorescent antibody ）technique, it was showed that strain was infectious. Purified virus was observed by Electron microscopic,we finded the average size of the isolated virus was 70-80 nm, rotundity ,without viral envelope. The result showed that isolated virus was porcine circovirus.