ISHS Acta Horticulturae 164: VI International Symposium on Virus Diseases of Ornamental Plants
DETECTION OF KALANCHOE LATENT VIRUS IN KALANCHOE BLOSSFELDIANA BY ENZYME-LINKED IMMUNOSORBENT ASSAY (ELISA) |
| Author: | S. S. Hearon |
| Abstract:
Double antibody sandwich ELISA was tested for use in screening naturally-infected Kalanchoë blossfeldiana for two strains of kalanchoë latent virus (KV-1 and KV-2). Gamma globulins from antisera produced in New Zealand white rabbits to each strain were partially purified by ammonium sulfate precipitation and conjugated with alkaline phosphatase. At monthly intervals from January to November, 2 g leaf samples from seven plants of cv. Golden Nugget and six plants of cv. Texas Sunset or healthy seedlings were ground in a semi-microblender in extraction buffer with polyvinylpyrollidone and tested in parallel tests against both conjugates. Flower samples were included in some tests. Plants of cv. Texas Sunset consistently reacted strongly with both conjugates, but the reaction with KV-2 antibody was stronger than that with KV-1 antibody. Plants of cv. Golden Nugget gave variable levels of reactions. Flowers and leaves gave comparable reactions. The threshold of reliable detection in these tests was estimated as 30 ng virus/ml buffer using dilutions of purified virus. In comparative ELISA tests using reagents prepared in this laboratory to KV-1 and KV-2 and commercially available kits for potato virus S detection in potatoes, the latter consistently detected KV-1 and KV-2 infected kalanchoës. | |
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