猪细小病毒细胞适应株的培育及鉴定  

Characterization of a porcine parvovirus cell culture-adapted strain

作　　者：张超范[1] 崔尚金[1] 戚亭[1] 陈长木[1] 苗岚飞[1] 谢红玲[1] 周盛华[1] 岳丰雄[1] 刘长明[1] 刘霓虹[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/猪传染病研究室,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2008年第5期362-366,共5页Chinese Journal of Preventive Veterinary Medicine

摘　　要：从临床表现为皮炎消瘦症状的仔猪肝脏中分离到1株病毒,经聚合酶链式反应(PCR)证实为猪细小病毒(PPV),采用仔猪原代肾细胞和传代ST细胞分离培养,经蚀斑克隆纯化,培育1株ST传代细胞培养适应毒株,命名为PPV-BQ2007株。免疫过氧化物酶单层细胞试验(IPMA)检测病毒抗原主要分布在细胞核及细胞质内。病毒感染细胞可被已知PPV阳性血清中和。免疫电镜可清晰见到聚集成团的大小不一的病毒粒子,近似圆形,无囊膜,直径大小约20nm～22nm。该分离株经ST细胞培养传代,能够产生典型的细胞病变,病毒滴度随代次显著增加,第30代后毒价达107TCID50/mL以上,且毒价和血凝价稳定。测序结果表明PPV-BQ2007株VP2基因与NCBI公布的皮炎型毒株Kresse株的同源性最高,达99.8%,在系统进化分支上处于同一个分支。PPV-BQ2007株传代细胞培养适应株的成功培育和鉴定,为进一步开展该病毒流行病学、致病机理、疫苗免疫与诊断研究等奠定了良好基础。A virulent strain of porcine parvovirus （PPV） was isolated from the liver of an infected piglet using the primary piglet kidney cell and the ST cells. A cell culture-adapted strain PPV-BQ2007 was obtained after a series of passages and three plaque purifications. The virus could induce typical cytopathogenic effects （CPE）, and the viral titer increased markedly after a series of passages and reached 10^7 TCID50/mL at the 30th passage. Immunoperoxidase monolayer assay （IPMA） showed that the virus antigens were distributed in the infected cell nucleolus and cytoplasm. The virus infectivity could be neutralized by the antisem against PPV. Partial sequence of VP2 gene was amplified by PCR and compared with eight PPV in GenBank, which showed 97.7 %-99.8 % nucleotide homology.

关 键 词：猪细小病毒 分离鉴定 毒株培育 

分 类 号：S852.659.2[农业科学—基础兽医学]