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Comparison of Mouse Monoclonal Antibodies and Polyclonal Antibodies of Chicken Egg Yolk and Rabbit for Assay of Carnation Etched Ring Virus. H. T. Hsu, American Type Culture Collection, 12301 Parklawn Drive, Rockville, MD 20852; R. H. Lawson, U.S. Department of Agriculture, Agricultural Research Service, Beltsville, MD 20705. Phytopathology 75:778-783. Accepted for publication 14 February 1985. 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, 1985. DOI: 10.1094/Phyto-75-778.

Serological detection of carnation etched ring virus (CERV) with mouse monoclonal antibodies, polyclonal rabbit antiserum, and chicken egg yolk antibodies was compared by ELISA (enzyme-linked immunosorbent assay) and IEM (immunoelectron microscopy). Hybridoma cells produced more antibodies in ascitic fluid in mice than in a culture medium. Sensitivity of CERV detection was similar in double antibody sandwich ELISA or triple antibody sandwich ELISA on PVC (polyvinyl chloride) plates coated with mouse ascitic fluid at a 1:1,000 dilution. Sensitivity of CERV detection was three- to four-fold lower on PVC plates coated with a 1:250 dilution of rabbit antiserum than with mouse monoclonal antibodies. Plates coated with antibodies from crude egg yolk or partially purified egg immunoglobulins failed to trap the virus. Rabbit antiserum-coated grids gave a linear increase in the number of virions attached to the grid over a concentration range of 3- 12 μg/ml. Treatment of grids with protein A before coating with antiserum increased the number of particles attached by five- to 10-fold. Only a few particles were attached at low virus concentrations without protein A treatment. In IEM on monoclonal antibody-coated grids, more particles were attached at higher virus concentrations when grids were treated with protein A, but there was not a linear increase in the number of particles with increasing virus concentration. Carnation etched ring virus particles treated with virus-specific monoclonal antibodies showed specific antibody attachment that was absent on virions treated with Prunus necrotic ringspot virus monoclonal antibody.