Articles
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
Article number
234_30
Pages
249 – 256
Language
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.
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.
Authors
S.-M. Wong, R.K. Horst
Keywords
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