A chemiluminescent immunoassay for the diagnosis of grapevine closteroviruses on nitrocellulose membrane.
A dot-immunobinding assay was adapted on enhanced chemiluminescence (DIBA-ECL), which employs luminol, a cyclic diacylhydrazide, as substrate for horseradish peroxidase conjugated with a secondary antibody, for the diagnosis of grapevine closteroviruses I and III. The sensitivity of DIBA-ECL was also compared to other immunoenzymatic methods. DIBA-ECL proved to be at least 16 times more sensitive than the dot-immunobinding assay using chloronaphthol/diaminobenzidine mixture as a substrate, which was at least twice as sensitive as DAS-ELISA, DAS-indirect avidin-biotin complex ELISA, and dot-immunobinding assay, using alkaline phosphatase as enzyme. Optimisation of all parameters involved in the DIBA-ECL procedure and its advantages are discussed.
Pollini
C P
CP
Institute of Plant Pathology, University of Bologna, Italy.
Giunchedi
L
L
Credi
R
R
D003101:Collodion; D005638:Fruit / Q000382:microbiology*; D007118:Immunoassay / Q000379:methods*; D015151:Immunoblotting / Q000379:methods*; D007124:Immunoenzyme Techniques; D008163:Luminescent Measurements; D008567:Membranes, Artificial; D010935:Plant Diseases; D010942:Plant Viruses / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
D008567:Membranes, Artificial; D003101:Collodion