【摘要】 为获得可用于犬细小病毒(CPV)感染的治疗和快速诊断的单克隆抗体,从病料中分离了1株犬细小病毒,经PCR扩增VP1全基因并进行序列分析,确定为CPV-2 a型。再将VP1基因克隆到真核表达质粒pc DNA3中,构建了基因疫苗pc DNA3-VP1。用该基因疫苗免疫BALB/c小鼠3次后,取抗体效价高的两只小鼠脾细胞与SP2/0细胞融合,经间接ELISA检测,结果获得了3株杂交瘤细胞株,该3株单抗与犬、猫细小病毒能发生反应,均不与犬瘟热病毒、流感病毒和犬腺病毒发生反应,诱生的腹水可体外中和犬细小病毒,其中两株单抗的中和效价为1∶256,1株单抗腹水的中和效价低于1∶128。结果表明,该CPV VP1基因疫苗可制备具有一定中和效价的CPV单克隆抗体。更多还原

【Abstract】 To prepare the monoclonal antibody( Mc Ab) against Canine parvovirus( CPV),a CPV-2 a was isolated and the VP1 gene was amplified by PCR and cloned into a eukaryotic expressing vector pc DNA3. The pc DNA3-VP1 was used as DNA vaccine and injected into Balb/c mice for three times. The spleen cells of two mice were fused with SP2/0 cells and three hybridomas were gotten by screening with indirect ELISA assay. The three monoclonal antibodies could react with CPV and FPV but could not react with CDV,AIV and CAV in vitro. The neutralization antibodies of two Mc Abs were proved to be 1: 256,and the other one was below 1: 128. The results suggest that the CPV VP1 gene vaccine can be used to prepare monoclonal antibody against CPV.更多还原