猪圆环病毒2型的分离与鉴定

Isolation and identification of porcine circovirus type 2

利用 PCR方法 ,从疑似断奶猪多系统衰竭综合征 (PMWS)的自然病例的淋巴结和脾脏中扩增出了预期长度的猪圆环病毒 2型 (Porcine Circovirus type 2 ,PCV 2 )的 DNA片段 ,并用限制性内切酶对 PCR产物进行了酶切鉴定。将经 PCR扩增和酶切鉴定为 PCV2阳性的组织样品匀浆液接种到无 PCV污染的 PK15细胞中传代 ,经间接免疫荧光检查 ,在接毒细胞内观察到了特异性免疫荧光 ;用接毒细胞制备超薄切片 ,在电镜下观察到了直径大约为 17nm的圆形病毒样粒子和大量不同形态的胞浆内包涵体。由此表明分离出的病毒为猪圆环病毒 2型。

By using PCR, The nucleic acid fragments of PCV2 were detected from the tissues of lymph node and spleen of clinical cases with PMWS. The PCR products were identified by restriction enzyme digestion. The PCV2 positive sample tissues were homogenated and inoculated to the PK15 cells that were not contaminated by PCVs. The PCV2 nucleic acid fragments were detected from the infected PK15 cell and the specific PCV2 antigen were shown through immunofluorescence in the infected cytoplasm. The viral particles and inclusion bodies were also discovered in the infected PK15 cells through electronic microscope. It can be concluded that the isolates are PCV2.