ISHS Acta Horticulturae 164: VI International Symposium on Virus Diseases of Ornamental Plants
DETECTION OF CARNATION ETCHED RING VIRUS USING MOUSE MONOCLONAL ANTIBODIES AND POLYCLONAL CHICKEN AND RABBIT ANTISERA. |
| Authors: | H.T. Hsu, R.H. Lawson |
| Abstract:
Serological detection of carnation etched ring virus (CERV) using mouse monoclonal antibodies and rabbit and chicken antisera were compared by ELISA and IEM. Hybridoma cells produced about 103 to 104 times higher titer of antibodies in ascitic fluid in mice than in cultured medium. Sensitivity of CERV detection was similar in double antibody sandwich ELISA or triple antibody sandwich ELISA in crude sap tests from infected leaf tissues or purified preparations on ELISA plates coated with mouse ascitic fluid diluted 1:1,000. Sensitivity of CERV detection was 3 to 4 fold lower on plates coated with 1/250 dilution of rabbit antiserum than with mouse monoclonal antibodies. Plates coated with antibodies from crude egg yolk or partially purified egg immunoglobulin failed to trap the virus. Purified CERV in the range of 1 to 14 μg/ml tested in IEM gave a linear response on rabbit antiserum coated grids. Pretreatment of grids with protein A increased the number of particles attached by 5 to 10 fold. Mouse antibody treated grids showed a higher concentration of attached virions with or without protein A pretreatment than grids treated with rabbit antiserum. | |
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