Enzyme linked immunosorbent assay for identification of whole Mycobacterium bovis isolates with the monoclonal antibodies

Objective: The development of a test system for identification of Mycobacterium bovis with the use of monoclonal antibodies (Mabs).Design: BALB/c mice were immunized with whole gamma-radiation-exposed M. bovis strains. The splenocytes of the immunized mice were used for producing hybridomes which generated Mabs specific to M. bovis. The specificity of the Mabs was determined with the ELISA procedure for the whole cells of the 11 mycobacteria of various species fixed in the wells of a microtitre plate. The optimal conditions for performing identification were selected.Results: Three Mabs specific to M. bovis have been produced which identify various epitopes of the protein with a molecular weight of 31 kD. The ELISA tests with the 2A1 Mab have been conducted for 213 strains of 15 species of mycobacteria. The technique has been shown to be highly specific and reproducible.Conclusion: An ELISA test system with a Mab for identification of M. bovis has been developed.