 Copyright The American Phytopathological Society |
Research. Development of Serological Procedures for Rapid, Sensitive, and Reliable Detection of Rice Grassy Stunt Virus. H. T. Hsu, Microbiologist, USDA, Agricultural Research Service, Beltsville Agricultural Research Center, Beltsville, MD 20705. H. Hibino, and P. Q. Cabauatan. Plant Virologist, and Senior Research Assistant, International Rice Research Institute, Manila, Philippines. Plant Dis. 74:695-698. Accepted for publication 6 April 1990. This article is in the public domain and not copyrightable. It may be freely reprinted with customary crediting of the source. The American Phytopathological Society, 1990. DOI: 10.1094/PD-74-0695. Hybridoma cell lines secreting IgG2a antibodies reactive to two strains of rice grassy stunt virus (RGSV) were established by the limiting dilution cloning technique. Serological detection of RGSV with mouse monoclonal antibodies and rabbit polyclonal antibodies was compared by enzyme-linked immunosorbent assay (ELISA), dot-blot immunoassay (DBI), and a latex agglutination test (LAT). A dilution endpoint of approximately 10–3 was obtained for both mouse monoclonal antibodies and rabbit polyclonal antiserum by LAT. The RGSV antigen in 2.5 ?l of a 10–3 dilution of infected sap could be detected by DBI when alkaline phosphatase-labelled antibodies were used. Using double antibody sandwich ELISA, an approximately 10-fold increase in sensitivity for RGSV detection was achieved. For practical purposes, however, the LAT provides the simplest procedure for routine detection of RGSV. |
