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Disease Detection and Losses

Detection of Lily Symptomless Virus by Immunodiffusion. Daina H. Simmonds, Department of Biology, University of New Brunswick, Fredericton, New Brunswick, Canada, E3B 5A3, Present address of senior author: Department of Biology, ELBA, Carleton University, Ottawa, Ontario, Canada K 1S 5B6; Bruce G. Cumming, Department of Biology, University of New Brunswick, Fredericton, New Brunswick, Canada, E3B 5A3. Phytopathology 69:1212-1215. Accepted for publication 22 May 1979. Copyright 1979 The American Phytopathological Society. DOI: 10.1094/Phyto-69-1212.

A simple immunodiffusion test for detecting lily symptomless virus (LSV) has been devised. LSV, a long flexuous rod (640 nm), does not readily diffuse through agar. Repeated freezing and thawing or sonication (4 min, 20 kc/sec) fragmented the virus sufficiently to allow diffusion through agar without destroying its antigenic properties. Because of its low concentration in sap, LSV was concentrated by isoelectric precipitation. LSV was precipitated from clarified sap at pH 4.0–4.4 with 1.0 M acetate buffer, pH 3.1. The precipitate was resuspended in 0.1 M phosphate buffer (pH 7.0) containing 0.05% Igepon T-77 and tested by the Ouchterlony double diffusion method. An LSV concentration of 1.8 μg/ml of clarified sap was detectable with this assay.