李痘病毒单克隆抗体及TAS-ELISA试剂盒的研制  

Preparation of the monoclonal antibody against Plum pox virus and TAS-ELISA kit

作　　者：马洁[1] 魏梅生[1] 粟智平 李桂芬[1] 

机构地区：[1]中国检验检疫科学研究院动植物检疫研究所,北京100121 [2]烟台出入境检验检疫局

出　　处：《植物检疫》2012年第2期9-12,共4页Plant Quarantine

基　　金：国家质检总局科技计划项目(2010IK264)

摘　　要：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。Plum pox virus was injected to BALB/c mice whose spleen cells were fused with myeloma cells SP2/0.Two hybridoma cell lines were obtained by the finite dilution and indirect ELISA.The 2 lines could secrete monoclonal antibody stably and were named as 3F1 and 7A8.The subtypes were identified as IgG1（3F1）and IgG3（7A8） by ELISA respectively.The titers of 3F1 and 7A8 ascetic fluids were 1：106 and 1：105 by indirect-ELISA respectively.A rabbit polyclonal antibody for capture and a monoclonal antibody as detection formed TAS-ELISA.The TAS-ELISA kit were demonstrated to react specifically to PPV-D and PPV-M,whereas the kit did not response to PVA,LMV,WMV-2,PVYN of the same genus.

关 键 词：李痘病毒 单克隆抗体 

分 类 号：S41-30[农业科学—植物保护]