水稻条纹病毒单克隆抗体的制备及检测应用

 用水稻条纹病毒(RSV)免疫的BALB/c小鼠脾细胞与SP2/0鼠骨髓瘤细胞融合,经筛选克隆,获得4株能稳定传代且分泌抗RSV单克隆抗体的杂交瘤细胞。各株单抗腹水的ELISA效价均在1:80000~1:5120000之间。Westernblot分析表明,4株单抗均与RSV的35kDa的外壳蛋白亚基有特异反应。建立了间接ELISA测定RSV的方法,4株单抗检测病汁液的稀释度能达到2560倍以上,与其它病毒无交叉反应。对江苏省部分县市的大田灰飞虱带毒率进行检测,结果显示灰飞虱的带毒率为12.5%~41.5%

Production of monoclonal antibodies to Rice stripe virus and application in virus detection

Four hybridoma cell lines secreting monoclonal antibodies (MAbs) against Rice stripe virus (RSV) were produced by fusing mouse myeloma cells (SP 2/0) with spleen cells from BALB/c immunized by the RSV particles. The titres of ascitic fluids of four MAbs ranged from 1:80000 to 1:5120000 tested by indirect ELISA. The result of Western blot showed that the four MAbs could react with the 35 kDa coat protein specifically. The four MAbs could detect RSV in plant sap with 1:2560 dilution by indirect ELISA successfully. The MAbs didn't cross-react with other tested plant viruses. Indirect ELISA was then used for detection of RSV in Laodelphax striatellus Fallen collected in Jiangsu Province. The proportion of the viruliferous Laodelphax striatellus ranged from 12.5% to 41.5%.