芜菁花叶病毒单克隆抗体的制备及检测应用  

Preparation of Monoclonal Antibodies to Turnip Mosaic Virus and Its Application for The Virus Detection

作　　者：施曼玲[1,2] 吴建祥[1] 郭维[1] 周雪平[1] 

机构地区：[1]浙江大学生物技术研究所 [2]杭州师范学院生命科学学院杭州310012

出　　处：《微生物学报》2004年第2期185-188,共4页Acta Microbiologica Sinica

基　　金：浙江省自然科学基金 ( 3 0 10 0 7) ;浙江省分析测试基金 ( 0 3 13 8)~~

摘　　要：先以芜菁花叶病毒 (TuMV)免疫BALB C小鼠 ,然后取其脾细胞使之与SP2 0鼠骨髓瘤细胞融合 ,经筛选、克隆 ,获得 4株能稳定传代并分泌抗TuMV单克隆抗体 (MAb)的杂交瘤细胞 ,并以之制备腹水单抗。 4株单克隆抗体腹水ELISA效价在 10 - 5～ 10 - 6 之间 ,仅对TuMV起特异性反应。Westernblot分析表明 ,4株单抗都能与TuMV34kD的外壳蛋白亚基起特异反应。利用TuMV的多抗兔血清和单抗腹水建立了三抗体夹心ELISA检测TuMV的方法 ,检测病叶的灵敏度为 1∶5 12 0倍 ,检测提纯TuMV病毒绝对量为 2 1 9ng。利用三抗体夹心ELISA测定出 7种作物上有TuMV侵染。Four hybridoma cell lines capable of secreting monoclonal antibodies (MAbs) against Turnip mosaic virus (TuMV) were obtained by fusing mouse myeloma cells (SP2/0) with spleen cells taken from the BAL B/C having been immunized by the TuMV particles. The four MAbs could specifically react with TuMV. The ELISA titres of ascitic fluids of four MAbs ranged from 10 -5 ～10 -6 . The result of Western blot showed that 4 MAbs can react specifically with the 34 kD coat protein submit of TuMV. TAS-ELISA was set up with polyclonal and MAbs for TuMV detection,and the MAbs could successfully detect 21.9ng purified TuMV or virus in plant sap at 1∶5120 dilution. TAS-ELISA were then used for detection of TuMV in seven crops.

关 键 词：芜菁花叶病毒 单克隆抗体 TUMV 寄主 杂交瘤细胞 田间作物 病原 检测方法 

分 类 号：S188[农业科学—农业基础科学]