COMPARISON OF ELISA AND IMMUNOBLOTTING TECHNIQUES FOR THE DETECTION OF CHERRY MOTTLE LEAF VIRUS

Formaldehyde treated cherry mottle leaf virus (ChMLV) and the isolated coat protein were used successfully for the production of polyclonal and monoclonal antibodies. The monoclonal antibodies had a titre of 1: 51200 and consisted of IgG1 and IgG2. The antibodies reacted with all 11 isolates of ChMLV, from five locations in Canada and the USA, inclu ded in this study. Several serological procedures were assessed to com pare their sensitivity for detecting ChMLV. Plate-trapped antigen ELIS A (PTA-ELISA) and dot-blot immunobinding assay (DBIA), using virus spe cific MAbs, were the most sensitive tests in this study. Triple antibo dy sandwich ELISA (TAS-ELISA) and Western blot were found to be less s ensitive. Dilution of the samples appeared to increase the sensitivity of both PTA-ELISA and Western blot detection. Young leaves and flower s of Prunus avium were the best tissue for detecting the virus which c ould also be detected in the fruit and leaves of P. tomentosa. April a nd May were optimal for detection of the virus in the field, whereas b oth April to May and August to September were optimal for screenhouse- grown plants.