CHEMILUMINESCENT AND COLORIGENIC DETECTION OF CHERRY LEAF ROLL VIRUS WITH DIGOXIGENIN-LABELED RNA PROBES

Digoxigenin-labeled RNA probes were used to detect cherry leaf roll vi rus in infected plants. A dot-blot hybridization immunoenzymatic assay in both crude sap extracts and partially purified tissue with a color igenic and chemiluminescent detection was developed. The use of the ne w AMPPD substrate was found to be effective in clarified sap extracts in conditions were the colorigenic detection method failed. Both detec tion assays were effective when using unfractionated nucleic acid prep arations, the chemiluminescent being five times more sensitive than th e colorigenic. The chemiluminescent hybridization assay makes it possi ble to detect the virus at the picogram level. The non-radioactive dot -blot hybridization techniques described here turned out to be very su itable for plant virus diagnosis. The sensitivity of this method and t hose obtained by ELISA or radioactive dot-blot described previously is compared.