Development of monoclonal-antibody-ELISA and -DIP-STICK immunoassays for Penicillium islandicum in rice grains
| Autor: | S. I. Phillips, R. A. Priestley, M. M. Macdonald, Frances M. Dewey |
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| Rok vydání: | 1990 |
| Předmět: |
Antigens
Fungal Hot Temperature food.ingredient medicine.drug_class Blotting Western Enzyme-Linked Immunosorbent Assay Biology Monoclonal antibody Immunofluorescence Microbiology Mice food medicine Animals Agar Immunoassay medicine.diagnostic_test Periodic Acid Penicillium Antibodies Monoclonal food and beverages Oryza Fungi imperfecti biology.organism_classification Molecular biology Penicillium islandicum Immunoglobulin G biology.protein Electrophoresis Polyacrylamide Gel Antibody |
| Zdroj: | Journal of General Microbiology. 136:753-760 |
| ISSN: | 0022-1287 |
| DOI: | 10.1099/00221287-136-4-753 |
| Popis: | Monoclonal antibodies (mAbs) were raised against Penicillium islandicum, a fungus commonly isolated from stored rice grains in South-East Asia. Mice were immunized by a direct, simple method; fresh cell-free surface washings from a solid agar slant culture were injected directly into the peritoneum without prior concentration. Hybridoma supernatants were screened by ELISA. Most of the mAbs raised cross-reacted with other storage fungi and/or uninfected rice grains but there were species-specific. One of these, PI01, was used to develop ELISA and DIP-STICK assays for the detection of P. islandicum in individual grains. All inoculated grains and approximately 90% of grains in natural infected samples from Indonesia tested positively, by ELISA, for P. islandicum. This result and those obtained for discoloured grains from both Indonesia and the Philippines, 32% and 14% respectively, are higher than those obtained by direct plating of surface-sterilized grains. Heat and periodate treatment of the PI01 antigen and binding on Western blots indicate that it is a glycoprotein of Mr greater than 90,000. Hyphae of all ages stained uniformly by immunofluorescence using the PI01 antibody but mature conidia stained only weakly. |
| Databáze: | OpenAIRE |
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