[建兰花叶病毒单克隆抗体的制备及其在兰花病毒检测中的应用]

[Production of monoclonal antibodies to Cymbidium mosaic virus and application in orchids virus detection].

作者信息

Meng Chun-Mei, Wu Jian-Xiang, Xie Li, Zheng Jin-Kai, Hong Jian

机构信息

Institute of Biotechnology, Zhejiang University, Hangzhou 310029, China.

出版信息

Wei Sheng Wu Xue Bao. 2007 Oct;47(5):928-31.

PMID:18062276

Abstract

Three hybridoma cell lines, 2C6, 5B7 and 12G9, secreting monoclonal antibodies (McAbs) against Cymbidium mosaic virus (CymMV) were produced by fusing mouse myeloma cells (SP2/0) with spleen cells from BALB/ C immunized by the CymMV particles. The three McAbs could specifically react with CymMV. The titres of ascitic fluids of two McAbs are up to 10(-6) in I-ELISA. Isotypes and subclasses of the the three McAbs belong to IgG1. Isotypes of light strains of the three McAbs all belong to kappa. They were used in antigen-coated plate (ACP)-ELISA for CymMV detection, and ACP-ELISA could successfully detect 0.487 ng of purified CymMV or virus in plant sap diluted 1:10240. The presence of CymMV in field Orchids tissues was investigated with ACP-ELISA.

摘要

通过将小鼠骨髓瘤细胞（SP2/0）与经建兰花叶病毒（CymMV）颗粒免疫的BALB/C小鼠脾细胞融合，制备了三株分泌抗建兰花叶病毒单克隆抗体（McAbs）的杂交瘤细胞系，分别为2C6、5B7和12G9。这三种单克隆抗体能与建兰花叶病毒发生特异性反应。在间接酶联免疫吸附测定（I-ELISA）中，两种单克隆抗体腹水的效价高达10^(-6)。这三种单克隆抗体的免疫球蛋白类型和亚类均属于IgG1。三种单克隆抗体轻链的免疫球蛋白类型均属于κ链。它们被用于包被抗原板（ACP）-ELISA检测建兰花叶病毒，ACP-ELISA能够成功检测到0.487 ng的纯化建兰花叶病毒或稀释1:10240的植物汁液中的病毒。利用ACP-ELISA对田间兰花组织中建兰花叶病毒的存在情况进行了调查。