Simple Latex Agglutination Test for Detecting Flexuous Rod-Shaped Viruses in Forage Legumes

Abstract

by Khan and Slack (13), except the latex Demski, J. W., Bays, D. C., and Khan, M. A. 1986. Simple latex agglutination test for detecting beads (Sigma, 10% w/v) were diluted to flexuous rod-shaped viruses in forage legumes. Plant Disease 70:777-779. about 0.6% (w/v). y-Globulin fractions (IgG) from crude serum were precipitated Flexuous rod-shaped viruses occurring in forage legumes were detected by a latex agglutination test with ammonium sulfate [equal volumes (LAT). This simple, sensitive, reliable, and rapid serological procedure detected alfalfa latent of saturated (NH 4)2SO 4 with antiserum (ALV), bean yellow mosaic (BYMV), clover yellow mosaic (CYMV), clover yellow vein (CYVV), diluted with nine volumes of distilled red clover vein mosaic (RCVMV), white clover mosaic (WCMV), and peanut mottle (PMV) viruses water]. Mixtures were stirred in a cold directly from leaf sap extracts of greenhouse and field samples of various clovers and peas. The room (4 Q overnight, then centrifuged at standard dilution procedure using purified virus preparations showed that LAT detected as little as ro m (4 C) o i n entifget 0.5, 0.5, 0.1, 0.2, 0.1, and 0.2 gg/ml of BYMV, CYVV, CYMV, RCVMV, WCMV, and ALV, 10,000 rpm for 0 min. Resulting pellets respectively. In crude sap extracts (1: 10, w/ v) from pea and clover, the detectable dilution end point were resuspended in 0.1 M tris-HCl for these viruses varied from 4 X 10-2 to4) 10-4.The capability of LAT to detect virus in composite buffer, pH 7.4 (to original volume of samples of infected and uninfected leaflets was 1 in 20 for BYMV and CYVV, 1 in 40 for ALV and antiserum), and dialyzed against the same RCVMV, and 1 in 100 for WCMV and CYMV. buffer to remove sulfate ions. Aliquots (5-ml) of several antiserum dilutions (e.g., 1:25, 1:50, 1:100, 1:200, 1:400, and 1:800) were mixed with equal volumes of latex Several viruses are known to infect of potato viruses (13). This study was beads and diluted 1:15 (v/v) in 0.85% forage legumes in the southeastern undertaken to determine the sensitivity NaCl. After incubating 1 hr at room United States. Some of these include and potential application of LAT in temperature, the mixture was centrifuged bean yellow mosaic virus (BYMV), clover forage legume virus research. for 30 min at 6,000 rpm (Sorvall SS-34