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Use of Monoclonal Antibodies to Characterize Grapevine Leafroll Associated Closteroviruses. J. S. Hu, Department of Plant Pathology, Cornell University, New York State Agricultural Extension Service, Geneva, NY, 14456; D. Gonsalves(2), D. Boscia(3), and S. Namba(4). (2)Department of Plant Pathology, Cornell University, New York State Agricultural Extension Service, Geneva, NY, 14456; (3)Centro di Studio del CNR sui Virus e le Virosi delle Colture Mediterranee, Bari, Italy, (4)Laboratory of Plant Pathology, University of Tokyo, Bunkyo-ku, Tokyo 113, Japan. Phytopathology 80:920-925. Accepted for publication 11 April 1990. Copyright 1990 The American Phytopathological Society. DOI: 10.1094/Phyto-80-920.

Stable hybridoma cell lines secreting monoclonal antibodies to the NY-1 isolate of grapevine leafroll associated closteroviruses (GLRaV) were produced by fusing spleen cells of immunized BALB/c mice and mouse myeloma cell line SP2/0-AG14. The monoclonal antibodies reacted with the NY-1 isolate and other type III isolates, but not with type I, II, and IV isolates. The reactions were the same in five different kinds of enzyme-linked immunosorbent assays (ELISA), immunosorbent electron microscopy, dot-immunoblotting, and Western blotting assays. Sensitivity of the monoclonal antibodies were very good for the detection of the virus in grape leaf tissue in double antibody sandwich ELISA. With the double gold labeling electron microscopy technique, we were able to detect serologically distinct particles of GLRaV in single leafroll affected grapevines. A sensitive Western blotting assay was developed to estimate the molecular weight of virus coat protein of the GLRaV from partially concentrated samples using the monoclonal antibodies.