适合猪圆环病毒2型敏感复制的PK15细胞株的克隆与鉴定

Cloning and identification of PK15 cells for the sensitive replication of porcine circovirus 2

为了有效提高猪圆环病毒2型(PCV2)在PK15细胞中增殖的效价,采用有限稀释法将PK15细胞进行克隆,通过间接免疫荧光方法筛选对PCV2敏感的克隆细胞。结果表明,通过克隆后获得1株对PCV2高度易感的细胞PK15-B1,该细胞接种PCV2后培养3d,病毒的TCID50达10-7.0/mL,病毒效价和生产速度明显高于PK15母细胞,且没有支原体、猪圆环病毒1型、牛黏膜病病毒、猪瘟病毒和脑心肌炎病毒等外源性微生物污染,为PCV2体外增殖和生物学特性的研究奠定了重要基础。

To improve the titers of porcine circovirus 2（PCV2）,PK15 cells were cloned using the limi-ting dilution method and the susceptible cells to PCV2 were selected by indirect immunofluorescence assay.In result,one clone,PK15-B1,could reliably produce PCV2 with high titers was selected from heterogeneous PK15 parental cells.The viral titer could be as high as 107.0TCID50/mL after the PK15-B1 cells inoculated with 100 TCID50 for 3 days.The levels of the viral titer and growth speed in PK15-B1 cells were significantly higher than those in PK15 parental cells.Moreover,the clone cells did not contaminated with porcine circovirus 1,bovine viral diarrhea virus,classical swine fever virus,encephalomyocarditis virus or mycoplasma.The results indicated that the PK15-B1 cell clone is reliably permissive to PCV2 replication and could be used to study the biological characterization of PCV2 and replication of PCV2 in vitro.