Method of detecting, isolating and purifying clostridium diffici

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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435 34, 435 39, 435803, 435842, 530413, 530825, 436520, 436534, 436829, G01N 33569, C12Q 104, C12Q 106

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active

048638528

ABSTRACT:
A method is provided for detecting the presence of C. difficile toxin A. Stool or other appropriate specimen is contacted with a reagent containing an available non-reducing galactose-alpha-1-3-galactosyl structure, which is a specific receptor for toxin A. The reagent may be intact cells, cell membranes, membrane fractions containing the toxin A receptor structure, glycoconjugates, as well as the purified toxin A receptor per se. Binding of toxin A is determined by conventional assay techniques. The method may also be used to isolate and purify toxin A. Conversely, immobilized toxin A is used to detect, isolate, or purify biological materials of interest containing the receptor structure.

REFERENCES:
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Spiro, R. et al., "Occurrence of .alpha.-D-Galactosyl Residues in the Thyroglobulins from Several Species", J. Biol. Chem., 259:9858-9866 (1984).
Wilkins et al., "Clostridial Toxins Active Locally in the Gastrointestinal Tract", Microbial Toxins and Diarrhoeal Disease, Pitman, London (Ciba Foundation Symposium 112), pp. 230-241 (Mar. 1985).
Lonroth et al., "Toxin A of Clostridium Difficile: Production, Purification and Effect in Mouse Intestine", Acta Path. Microbiol. Immunol. Scand. Sect B, 91: 395-400 (1983).
Lyerly, et al., "Enzyme-Linked Immunosorbent Assay for Difficile Toxin A", J. Clin. Microbial., 17: 72-78 (1983).
Thelestram, M. et al., "Cytopathogenic Action of Clostridium Difficile Toxins", J. Toxicol.-Toxin Reviews, 3: 139-180 (1984).

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