VARIANTS AND DISTINGUISHING FEATURES OF ENZYME IMMUNOASSAY OF PLANT VIRUSES

Enzyme – linked immunosorbent assay (ELISA) is widely used for detecting plant viruses in practice.
We have prepared kits for the detection of 10 plant viruses, mainly viruses of ornamental plants (carnation, chrysanthemum, freesia, lily). We have obtained reactives (virus antigens, antibodies, conjugates antibody-enzymes et al.) and worked on standartization of assay procedure.
We were investigating influence of some factors on sensitivity of virus assay: comparision of label-enzyme (alkaline phosphatase or horse-radish peroxidase), substrates of enzymes, quality of antibodies and so on.
By using the classical method of ELISA sandwich, we get the detection limit 1–10 ng per ml.

We have investigated the detection of plant viruses by some other methods and variants: 1) direct ELISA by using anti-species antibodies; 2) ELISA with biotinylated antibodies compared to enzyme-labeled antibodies; 3) dot-blot immunoassay on nitrocellulose or nylon membranes; 4) immunoblotting.
By using avidin-biotin system we have tested following reactives and variants: conjugates avidin – enzymes; the bridge method-biotinylated antibodies (avidin) biotinylated enzyme; preformed complex of avidin or streptavidin; biotinylated polyenzyme.
The detection limits, advantages and limitations of the above are discussed.

VIII International Symposium on Virus Diseases of Ornamental Plants

A. Zeidaka, L. Dzirkale, O. Skrodelis, J. Ignass

https://doi.org/10.17660/ActaHortic.1994.377.21