南方水稻黑条矮缩病毒和水稻黑条矮缩病毒的单抗制备及其检测应用

 用与牛血清白蛋白偶联的南方水稻黑条矮缩病毒（Southern rice black-streaked dwarf virus,SRBSDV）衣壳蛋白的C端12个氨基酸多肽为抗原免疫BALB/c小鼠,经细胞融合、筛选、克隆,获得2株能稳定传代并分泌抗SRBSDV和水稻黑条矮缩病毒（Rice black-streaked dwarf virus,RBSDV）单克隆抗体（MAb）的杂交瘤细胞株3F1、5G1。3F1、5G1单克隆抗体腹水间接ELISA效价达10^-6,抗体类型及亚类均为IgG1, kappa链。 Western blot分析表明,2株单克隆抗体均与SRBSDV和RBSDV的外壳蛋白亚基有特异反应。利用单克隆抗体3F1建立的dot-ELISA检测方法能准确、特异、灵敏地检测田间稻飞虱及水稻样品中的SRBSDV和RBSDV。SRBSDV和RBSDV单克隆抗体的制备及检测方法的建立为水稻黑条矮缩病的诊断、预测预报及科学防控提供了技术支撑。

Development of monoclonal antibodies against Southern rice black-streaked dwarf virus and Rice black-streaked dwarf virus and their application in virus detection

Two hybridoma cell lines (3F1 and 5G1) secreting monoclonal antibody (MAb) against Southern rice black-streaked dwarf virus (SRBSDV) and Rice black-streaked dwarf virus (RBSDV) were produced by fusing mouse myeloma cells (SP2/0) with spleen cells from BALB/c mouse immunized by a polypeptide containing 12 amino acids from the C- terminus of SRBSDV coat protein coupled with bovine serum albumin (BSA). The indirect-ELISA titres of ascitic fluids of the two MAbs were 10^-6. Both the MAbs had IgG1 isotypes with κ light chain. Western blot analysis indicated that both the MAbs could specifically react with the coat protein of SRBSDV and RBSDV. Based on the produced MAb, a dot-ELISA was established for detection of SRBSDV and RBSDV in rice planthoppers and rice samples. The detection method based on the produced anti-SRBSDV and RBSDV MAb provided technology for diagnosis, forecast and control of rice black-streaked dwarf disease.