Date of Award
Doctor of Philosophy
A direct sandwich solid-phase immunoassay, using antibody-coated polystyrene beads, was developed for the detection and quantitation of soybean mosaic virus (SMV) in seeds. Virus-specific IgG was passively adsorbed to the solid phase, and virus antigen bound by the immobilized antibody was demonstrated by subsequent binding of anti-SMV IgG labelled with either ('3)H (for SPRIA) or alkaline phosphatase (for ELISA). Optimal conditions for coating the solid phase, specificity of the immune reaction, and other kinetics and sensitivities of the assay were also defined for the purpose of standardizing the assay. A sensitivity level of 5 to 25 ng of purified SMV/ml was obtained, depending on the nature of the label used. The addition of healthy seed extracts did not interfere with the assay's ability to detect purified virus, indicating its potential for seed analysis. The assay easily distinguished infected from healthy seed extracts, as well as identifying various levels of infection;Technical simplicity, reproducibility and versatility of the solid-phase procedure make it diagnostically useful for detecting SMV in individual and bulked seeds.
Digital Repository @ Iowa State University, http://lib.dr.iastate.edu/
Glenda Rae Bryant
Bryant, Glenda Rae, "The development of serodiagnostic procedures for the detection and quantitation of soybean mosaic virus in seeds " (1981). Retrospective Theses and Dissertations. 6894.