---

作 者 ：刘欢,倪跃群,饶黎霞,吴建祥,周雪平

期 刊 ：植物病理学报 2013年 43卷 1期 页码：27-34

关键词：南方水稻黑条矮缩病毒；水稻黑条矮缩病毒；多肽；单克隆抗体；dot-ELISA；

Keywords:Southern rice black-streaked dwarf virus, Rice black-streaked dwarf virus, polypeptide, monoclonal antibody, dot-ELISA,

摘 要 ：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。

Abstract: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.

---