Development and application of a monoclonal antibody against Thiothrix spp
| Autor: | Stephen G. Zam, Bonnie O'Brien, Gabriel Bitton, R. L. Brigmon |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
medicine.drug_class
Carbohydrates Fluorescent Antibody Technique Enzyme-Linked Immunosorbent Assay Monoclonal antibody Immunofluorescent microscopy Applied Microbiology and Biotechnology Epitope Microbiology Microbial ecology medicine Environmental Microbiology Thiothrix Ecology biology Antibodies Monoclonal biology.organism_classification Isolation (microbiology) Thiobacillus Molecular biology Antibodies Bacterial biology.protein Antibody Bacteria Food Science Biotechnology Research Article |
| Zdroj: | Applied and environmental microbiology. 61(1) |
| ISSN: | 0099-2240 |
| Popis: | Historically, methods used to identify Thiothrix spp. in environmental samples have been inadequate because isolation and identification procedures are time-consuming and often fail to separate Thiothrix spp. from other filamentous microorganisms. We described a monoclonal antibody-based enzyme-linked immunosorbent assay (ELISA) procedure which was used to identify Thiothrix spp. in wastewater, artesian springs, groundwater, and underwater subterranean samples. The ELISA utilized monoclonal antibody T3511 to a species-specific carbohydrate epitope of Thiothrix spp. No cross-reactions were observed among non-Thiothrix strains consisting of 12 species and nine genera. In field trials, the ELISA identified 100% of 20 biochemically and cytologically confirmed Thiothrix spp.-containing samples with no false positives. Indirect immunofluorescent microscopy utilizing T3511 was effective for wastewater samples but not for those from natural spring water because of background fluorescence in the latter. In addition, electron micrographs of Thiothrix spp. labeled with T3511-biotin-anti-mouse antibody-gold showed that epitope T3511 was intracellular both in laboratory strains and environmental isolates. The minimum level of detection of the ELISA was 0.10 microgram/ml. |
| Databáze: | OpenAIRE |
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