Use of tick cell culture-derived Anaplasma marginale antigen in a competitive ELISA for serodiagnosis of anaplasmosis

Anaplasma marginale was propagated in a continuous tick cell line and detergent-solubilized infected cells were used as antigen in a competitive ELISA (C-ELISA) for detection of Anaplasma-specific antibody in bovine sera. Positive control sera competed well (greater than or equal to 35% inhibition) with an A. marginale-specific monoclonal antibody for binding to this antigen, while negative sera failed to compete (<35% inhibition). The C-ELISA was compared to the standard complement-fixation test (CPT) using 2,208 bovine sera. Overall, C-ELISA was more sensitive than CFT (24.9% versus 9.4%), mainly because CFT yielded "suspicious" or "anti-complementary" results in 10.5% of the sera and also failed to identify several vaccinated and carrier cattle that were C-ELISA-positive. The apparent agreement between CFT and C-ELISA was 89.6% and the kappa value was 0.6. These results show that this C-ELISA would be a suitable replacement of the CFT as the standard test for detection of A. marginale antibody.