Detection of Lily Symptomless Virus by Immunodiffusion

Abstract

Lily symptomless virus (LSV), a long flexuous rod (640 nm), does not readily diffuse through agar. Repeated freezing and thawing or sonication (4 min, 20 kc/s) 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 [of Lilium] 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 .mu.g/ml of clarified sap was detectable with this assay.