百合无症病毒单克隆抗体的制备及检测应用  

Production of monoclonal antibodies to Lily symptomless virus and application in lily virus detection

作　　者：吴建祥[1] 刘成科[1] 洪健[1] 刘文洪 叶美琴[3] 

机构地区：[1]浙江大学生物技术研究所,杭州310029 [2]浙江中医学院,杭州310053 [3]浙江丽水市农业局,丽水323000

出　　处：《微生物学报》2005年第4期580-583,共4页Acta Microbiologica Sinica

基　　金：浙江省科技厅资助项目(2002C32128);国家科技部农业科技成果转化专项资助项目(02EFN213300269)~~

摘　　要：用百合无症病毒(Lilysymptomlessvirus,LSV)免疫的BALBC鼠脾细胞与SP20鼠骨髓瘤细胞融合,经筛选克隆,获得4株能稳定传代并分泌抗LSV单克隆抗体(MAb)的杂交瘤细胞(2A2、5H9、5H2和5E12),并分别制备它们的单抗腹水。4株单克隆抗体腹水间接ELISA效价达10-6,5H9和5E12的抗体类型及亚类均为IgG1,而2A2和5H2均为IgG3,4株单克隆抗体的轻链均为κ链。利用单克隆抗体建立了抗原包被间接ELISA(ACP-ELISA)检测LSV的方法。病叶作1300倍稀释、提纯LSV病毒浓度为18ngmL(每孔的病毒绝对量为1.8ng)时,该方法仍能检测到病毒。利用ACP-ELISA检测了田间样品,发现LSV在百合上发病很普遍。Four hybridoma cell lines, 2A2, 5H9, 5H2 and 5E12, secreting monoclonal antibodies (MAbs) against Lily symptomless virus (LSV) were produced by fusing mouse myeloma cells (SP2/0) with spleen cells from BALB/C immunized by the LSV particles. The four MAbs could specifically react with LSV. The titres of ascitic fluids of the four MAbs are up to 10 -6 in ELISA. Isotypes and subclasses of 5H9 and 5E12 belong to IgG1 while those of 2A2 and 5H2 belong to IgG3.Isotypes of light strains of the four MAbs all belong to κ. The four MAbs were used in antigen-coated plate (ACP)-ELISA for LSV detection, and ACP-ELISA could successfully detect 1.8 ng of purified LSV or virus in plant sap diluted 1:300.The presence of LSV in field lily tissues was investigated with ACP-ELISA.

关 键 词：百合无症病毒 单克隆抗体 ACP-ELISA 

分 类 号：S436.8[农业科学—农业昆虫与害虫防治]