The investigation aimed at preparing an ELISA kit for examination of bovine sera for brucellosis and evaluating the diagnostic properties of the test. Lipopolysaccharide extract (LPS) was used as the antigen for microplate coating, antibodies against IgG of bovine sera with horseradish peroxidase were used as the conjugate and ABTS with H202 as the substrate. A weak-positive serum prepared from the second International Standard of anti-Brucella abortus Serum (ISaBaS II) and a negative serum obtained by mixing sera from Brucella-free cows were used as controls. Evaluation of the diagnostic usefulness of the ELISA was performed by comparison of the results obtained with those in the rose bengal test (RBT) and complement fixation test (CFT). The examinations involved 212 bovine sera positive in the RBT, 60 sera positive in the CFT, 5 standard sera, and 1005 sera negative in the RBT. A high correlation was found between the ELISA and CFT results (55 sera out 60 sera) and no correlation between the ELISA and RBT results. Only 60 out of 212 sera positive in RBT gave positive results in the ELISA. All standard sera were positive in the ELISA and only 1 serum from those negative in the RBT was doubtful in the ELISA. The ELISA proved to be a suitable method for diagnosis of brucellosis in cattle.
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