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将纯化的李痘病毒(Plum pox virus,PPV)制剂免疫BALB/c小鼠,用SP2/0骨髓瘤细胞与经李痘病毒免疫的BALB/c小鼠的脾细胞融合,有限稀释法克隆和间接ELISA法筛选出2株稳定分泌李痘病毒单克隆抗体的杂交瘤细胞株3F1,7A8。用间接ELISA方法对所获得的2个杂交瘤细胞株进行亚型鉴定分别为IgG1、IgG3。间接ELISA方法测定腹水效价分别为3F1:1.0×106,7A8:1.0×105。以多克隆抗体为包被抗体、单克隆抗体为检测抗体的TAS-ELISA试剂盒与李痘病毒的D株系、M株系的病毒分离物均有反应,与同属的马铃薯A病毒、莴苣花叶病毒、西瓜花叶病毒2号、马铃薯Y病毒坏死株系不发生交叉反应。
BALB / c mice were immunized with the purified preparation of Plum pox virus (PPV), fused with spleen cells of SP2 / 0 myeloma cells and BALB / c mice vaccinated with Leepox virus, and cloned by limiting dilution Two hybridoma cell lines, 3F1 and 7A8, secreting monoclonal antibodies against the pox virus were screened by indirect ELISA. The subtype of two hybridoma cell lines obtained by indirect ELISA was IgG1 and IgG3 respectively. Indirect ELISA method for the determination of ascites titers were 3F1: 1.0 × 106,7A8: 1.0 × 105. Using polyclonal antibody as coated antibody, TAS-ELISA kit with monoclonal antibody as detection antibody reacted with virus of D strain and M strain of plum pox virus, and tested with the same potato A virus, lettuce flower Leaf virus, watermelon mosaic virus 2, potato Y virus necrosis strains do not cross-react.