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Serological Detection of Seed-Borne Barley Stripe Mosaic Virus by a Simplified Radial-Diffusion Technique. S. A. Slack, Department of Plant Pathology, University of California, Davis 95616, Current address of senior author: Department of Plant Pathology, University of Wisconsin, Madison 53706; R. J. Shepherd, Department of Plant Pathology, University of California, Davis 95616. Phytopathology 65:948-955. Accepted for publication 26 March 1975. DOI: 10.1094/Phyto-65-948.

A simplified radial-diffusion test which eliminated tissue grinding, well cutting, and chemical treatment of the antigen prior to exposure to antibody was developed for the serological detection of barley stripe mosaic virus. Immunospecific reactions were assessed 24-36 hours after embedding leaf tissue directly into a 0.5% agar matrix containing antiserum and an appropriate virus dissociating reagent. Approximately 500-600 specimens could be assayed in a 90-mm petri plate at the rate of 100-200 samples per person per hour. This radial-diffusion test could detect about 1 µg BSMV per ml, approximately a 10-fold greater sensitivity than conventional double-diffusion tests. Assay sensitivity was affected by the choice of dissociating agent and the aggregation state of BSMV injected into rabbits. In general, 0.5% Leonil SA detergent and antiserum against dissociated BSMV (D-protein) in the agar matrix gave the best results. It is proposed that BSMV D-protein antiserum should be used for immunodiffusion-in-gel serodiagnosis of seed-borne BSMV.

Additional keywords: mass testing, formalinized protein.