ELISA sanduíche para detecção de toxina beta produzida pelo Clostridium perfringens tipos B e C

Abstract

An ELISA test was developed and standardized to detect beta toxin produced by the Clostridium perfringens types B and C. The beta toxin was purified by metal affinity chromatography and immunoaffinity chromatography and detoxified by the method of controlled iodination. Anti-toxin IgG raised in sheep were purified on a beta toxin affinity column and conjugated to horse radish peroxidase. The sandwich ELISA detected 2.44ng/ml of the purified beta toxin in PBS buffer and intestinal content. The ELISA compared favourably to the toxicity test in mice, showing an 1000 fold higher sensitivity, detecting 0,01 DMM/mL, of nonpurified beta toxin in intestine content artificially contaminated.