ISHS Acta Horticulturae 234: VII International Symposium on Virus Diseases of Ornamental Plants
COMPARISON OF ANTIGEN AND ANTIBODY-COATED ENZYME-LINKED IMMUNOSORBENT ASSAY PROCEDURES FOR THE DETECTION OF THREE ISOLATES OF PURIFIED APPLE MOSAIC VIRUS OR PRUNUS NECROTIC RINGSPOT VIRUS |
| Authors: | S.-M. Wong, R.K. Horst |
| Abstract:
The objectives of this study were: 1) to establish an ELISA system to detect apple mosaic virus and prunus necrotic ringspot virus in rose, and 2) to compare the sensitivity of antigen and antibody-coated ELISA procedures. Attempts to develop antigen-coated ELISA to detect viruses in crude extracts were unsuccessful; however, an antibody-coated ELISA method using 1 ug coating antibody and 1:500 dilution of enzyme-antibody conjugate (1 mg/ml) gave highly specific reactions. ELISA test plates with added substrate developed no non-specific background readings when left overnight at room temperature. Dilution endpoints of 6 to 54 ng were obtained with three isolates of purified apple mosaic virus or prunus necrotic ringspot virus in antibody-coated ELISA. The occurrence of extremely high dilution endpoints of 0.1 to 0.5 ng with purified apple mosaic virus or prunus necrotic ringspot virus in antigen-coated ELISA but not in the presence of crude extracts, and the failure of dilution curves to cluster suggested competition of host protein and virus coat protein for binding sites. | |
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