Detection of antibodies to Cowdria ruminantium in the serum of domestic ruminants by indirect ELISA

Abstract

A solid phase enzyme immunoassay for the detection of antibodies to Cowdria ruminantium in the serum of domestic ruminants was developed by using microorganisms cultivated on bovine umbilical endothelial cells as antigen. When the culture showed 90 % lysis, the supernatant was centrifuged, sonicated and coated on polystyrene microtiter plates. Antibodies were detected as early as 9 days after experimental immunization of goats. The sensitivity of the ELISA calculated with 73 ruminant sera ranged between 97.3 % and 98.6 %.The overall specificity of the test was 97 % (N = 2925). However, the specificity was far lower for sheep (94.4 %, N = 881) than for goats (98.6 %, N = 651) and cattle (97.8 %, N = 1393). Crossreactivity which could explain some of the false positive reactions, was found between Cowdria antigen and sera raised against Ehrlichia bovis (1 bovine positive out of 2 inoculated) or E. ovina (2 sheep out of 2 inoculated became positive) but not with E. phagocytophila. The intra-assay and inter-assay variability were 7.5 % and 7.8 % respectively, indicating a good reproducibility of the ELISA.