猪圆环病毒河南株的分离与鉴定

Isolation and Identification of Porcine Circovirus Henan Strain

[目的]为研究猪圆环病毒的分子生物学特征和发病机理奠定基础。[方法]通过无菌操作采集病猪的脾脏和淋巴结,经胰蛋白酶消化处理后接种PK-15细胞,对病毒进行分离和纯化,并通过间接免疫荧光试验对其进行检测。[结果]断奶期的发病猪被毛粗糙、消瘦、精神沉郁、食欲不振,以5～10周龄猪的症状最明显。猪生长发育明显受阻,甚至导致死亡。从病料中和细胞中扩增出约540bp特异性片段。淋巴结、脾组织细胞核内均有大量无囊膜的病毒粒子,直径约17nm,呈球状,符合PCV病毒粒子特征。第6代细胞病毒液经间接免疫荧光检测可见PCV特征的荧光斑,而胞浆中无荧光斑,表明毒株有感染性。[结论]该试验中分离的病毒被鉴定为猪圆环病毒。

[Objective] The research aimed to lay the foundation for studying the molecular biological characteristics and pathogenesis of porcine circovirus.[Method] The spleen and lymph nodes of the diseased pigs collected by aseptic operation were treated digesting with trypsinase and inoculated on PK15 cells to isolate and purify the virus.And it was detected in the indirect immunofluorescence assay.[Result] The diseased pigs in the weaning period became thin and depressed with rough hair and inappetence,and the most obvious symptom was in 5～10 weeks old pigs.The growth and development of the pigs obviously suffocated and this even led to death.About 540 bp specific fragments were amplified from the diseased materials and cells.There were a large number of virions without envelope membrane in the nucleus of lymph nodes and spleen tissues,their diameter was about 17 nm and they showed sphericity,which accorded with the characeristics of PCV virion.Fluorescent spot characteristic of PCV could be seen in the virus liquid of the 6th-generation cells,while there was no fluorescent spot in the cytoplasm,which showed that the virus strain was infectious. [Conclusion] The isolated virus in the test was identified as porcine circovirus.