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 virus in infected plants. A dot-blot hybridization immunoenzymatic assay in both crude sap extracts and partially purified tissue with a colorigenic and chemiluminescent detection was developed. The use of the new AMPPD substrate was found to be effective in clarified sap extracts in conditions were the colorigenic detection method failed. Both detection assays were effective when using unfractionated nucleic acid preparations, the chemiluminescent being five times more sensitive than the colorigenic. The chemiluminescent hybridization assay makes it possible to detect the virus at the picogram level. The non-radioactive dot-blot hybridization techniques described here turned out to be very suitable for plant virus diagnosis. The sensitivity of this method and those obtained by ELISA or radioactive dot-blot described previously is compared.
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CEBAS (CSIC), Murcia, Spain.
Sánchez-Navarro
J A
JA
Sánchez-Pina
M A
MA
Pallás
V
V
D015152:Blotting, Northern; D004076:Digoxigenin; D005638:Fruit / Q000382:microbiology*; D015151:Immunoblotting / Q000379:methods*; D008163:Luminescent Measurements; D017860:Nepovirus / Q000235:genetics / Q000302:isolation & purification*; D015347:RNA Probes; D012367:RNA, Viral / Q000235:genetics*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods*
D015347:RNA Probes; D012367:RNA, Viral; D004076:Digoxigenin