Production of monoclonal antibodies to Cymbidium mosaic virus and application in Orchids virus detection

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 κ. They were used in antigen-coated plate (ACP) -ELISA for CymMV detection, and ACP-ELISA could successfully detect 0.487ng of purified CymMV or virus in plant sap diluted 1∶10240. The presence of CymMV in field Orchids tissues was investigated with ACP-ELISA.

Key words:Cymbidium mosaic virus; monoclonal antibodies; ACP-ELISA