Development of monoclonal-antibody-ELISA, -DOT-BLOT and -DIP-STICK immunoassays for Humicola lanuginosa in rice
| Autor: | Frances M. Dewey, M. M. Macdonald, S. I. Phillips |
|---|---|
| Rok vydání: | 1989 |
| Předmět: |
Immunogen
food.ingredient Antigens Fungal medicine.drug_class Dot blot Food Contamination Monoclonal antibody Microbiology food Antigen medicine Agar Immunoassay biology medicine.diagnostic_test food and beverages Antibodies Monoclonal Oryza Fungi imperfecti biology.organism_classification biology.protein Food Microbiology Microscopy Electron Scanning Mitosporic Fungi Antibody |
| Zdroj: | Journal of general microbiology. 135(Pt 2) |
| ISSN: | 0022-1287 |
| Popis: | Monoclonal antibodies (MAbs) were raised against Humicola lanuginosa, a thermophilic, saprophytic fungus that is commonly isolated from freshly harvested rice grains in Indonesia. Mice were immunized by direct injection into the peritoneum, without prior concentration, of fresh cell-free surface washings from a solid agar slant culture. Hybridoma supernatants were screened by ELISA using wells coated with a dilution of the immunogen. From one fusion 403 hybridoma clones were obtained yielding 52 cell lines secreting antibodies positive for H. lanuginosa. Twelve cell lines were re-cloned, grown in bulk and tested against other storage fungi. Most of the MAbs raised were IgM antibodies that cross-reacted with several of the storage fungi and/or uninfected rice grains. However, the IgM antibody EC6 did not recognize antigens from rice grains and cross-reacted strongly with only one other test fungus, Penicillium variabile, and partially with two others. This MAb was used to develop a highly sensitive DIP-STICK immunoassay to detect the fungus in infected grains. These assays are simple to perform, require no equipment and are suitable for field use by untrained workers. |
| Databáze: | OpenAIRE |
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