西瓜花叶病毒单克隆抗体及TAS-ELISA试剂盒的研制  

Preparation of the monoclonal antibody against Watermelon mosaic virus and TAS-ELISA kit

作　　者：梁新苗[1] 李桂芬[2] 马洁[2] 邓丛良[1] 边勇[1] 张永江[2] 李建光[1] 骆卫峰[1] 周琦[1] 

机构地区：[1]北京出入境检验检疫局,北京101312 [2]中国检验检疫科学研究院动植物检疫研究所

出　　处：《植物检疫》2013年第2期53-56,共4页Plant Quarantine

基　　金：国家质检总局科技计划项目(2009IK249)

摘　　要：将纯化的西瓜花叶病毒(Watermelon mosaic virus,WMV)制剂免疫BALB/c小鼠,用SP2/0骨髓瘤细胞与经西瓜花叶病毒免疫的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1株稳定分泌西瓜花叶病毒单克隆抗体的杂交瘤细胞株6C6。用间接ELISA方法对所获得的杂交瘤细胞株进行亚型鉴定为IgG1。间接ELISA方法测定腹水效价为1∶105。以单克隆抗体为包被抗体、多克隆抗体为检测抗体的TAS-ELISA试剂盒与引自ATCC的西瓜花叶病毒毒源PV-27、PV-379、PV-394、PV-511分离物均有反应,与同属的马铃薯A病毒(Potato virus A)、莴苣花叶病毒(Lettuce mosaicvirus virus)、李痘病毒(Plum pox virus)不发生交叉反应,与同属的番木瓜环斑病毒呈弱阳性反应。。Watermelon mosaic virus was injected to BALB/c mice whose spleen cells were fused with myeloma cells SP2/0.One hybridoma cell line was obtained by the finite dilution and indirect ELISA.The line could secrete monoclonal antibody stably and were named as 6C6.The subtype was identified as IgG1 by ELISA.The titer of 6C6 ascitic fluids was 1:105 by indirect-ELISA.A monoclonal antibody for capture and a rabbit polyclonal antibody as detection formed TAS-ELISA.The TAS-ELISA kit were demonstrated to react specifically to Watermelon mosaic virus,whereas the kit did not response to PVA(potato virus A),LMV(Lettuce mosaic virus),PPV(Plum pox virus) of the same genus,but weakly responsed to PRSV(papaya ringspot virus) of the same genus.

关 键 词：西瓜花叶病毒 单克隆抗体 TAS-ELISA试剂盒 

分 类 号：S41-30[农业科学—植物保护]