Development and evaluation of various enzyme-linked immunosorbent assays for the detection of Clostridium perfringens β anti-toxins

The aim of our work was to develop an enzyme-linked immunosorbent assay for the detection of antibodies against the Clostridium perfringens beta toxin. For this purpose, five different ways of performing an enzyme-linked immunosorbent assay were investigated. Positive and negative sera of different animals and partially purified beta toxin were used. In all enzyme-linked immunosorbent assay tests, microplates were first coated with monoclonal antibodies against the C. perfringens beta toxin. Actually, the first three ways of performing enzyme-linked immunosorbent assay proved to be an inhibition or a blocking enzyme-linked immunosorbent assay. In the first of these modifications, the examined serum was added on a microplate after the toxin. In the second two tests, they were added simultaneously after they were incubated together (60 min at room temperature or overnight at 4 degrees C, respectively). An anti-toxin conjugate was used for the detection. It was also used in a competitive enzyme-linked immunosorbent assay, where it was added together with the examined serum on the microplate, to which the toxin was already bound. The fifth way of performing an enzyme-linked immunosorbent assay differed from others by the use of conjugated anti-species immunoglobulin for the detection. The biggest differences in absorbances between positive and negative sera were found in the blocking enzyme-linked immunosorbent assay, where the mixture of the toxin and the examined serum were previously incubated overnight at 4 degrees C. The smallest differences in absorbance were found when anti-species conjugates were used. (C) 1999 Federation of European Microbiological Societies. Published by Elsevier Science B.V. All rights reserved.