Development of monoclonal antibody‐based immunological assays for the detection of live propagules of Rhizoctonia solani in soil

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Abstract
Mouse monoclonal antibodies (mAbs) and rabbit (polyclonal) antiserum were used to develop DIAGNOSTIC‐ELISA, double‐antibody‐sandwich‐ELISA (DAS‐ELISA), DIP‐STICK and immuno‐fluorescence colony staining immunoassays for the specific detection of Rhizoctonia solani in soil. mAbs were raised against an anastomosis group 4 isolate of R. solani. Mice were immunized using either phosphate‐buffered saline (PBS) suspensions of lyophilized mycelium plus Quil A adjuvant, or with a solubilized acetone precipitate prepared from cell‐free surface washings from solid slant cultures. Polyclonal antisera were raised in rabbits using PBS suspensions of lyophilized mycelium and Quil A adjuvant. Hybridoma supernatants and rabbit antisera were screened by ELISA. Four of the cell lines raised produced mAbs that were species‐specific. They recognized antigens from R. solani by ELISA and immunofluorescence, but not other related or unrelated species of soil‐borne fungi. The remaining cell line produced mAbs that cross‐reacted slightly, by ELISA, with antigens from R. cerealis. These mAbs did not recognize R. cerealis by immunofluorescence, or other related or unrelated soil‐borne fungi, by ELISA.