A sensitive ELISA technique for the diagnosis of Anaplasma marginale infections

A sensitive enzyme-linked immunosorbent assay (ELISA) technique using a horse radish peroxidase conjugate is described for measuring Anaplasma marginale antibodies in bovine serum. This technique utilizes two antigen preparations, a 'negative' antigen derived from an animal prior to infection and a 'positive' antigen derived from A. marginale-infected red cells from the same animal following infection. This markedly reduces cross-reactions which are a result of isoantigens. Absorbance values obtained using the 'negative' antigen are subtracted from those obtained using the 'positive' antigen to give a net figure. Of 100 A. marginale-positive sera tested no false negative results were obtained. All 11 animals maintained tick-free after initial diagnosis of naturally transmitted anaplasmosis were positive 3 years later, 15 A. marginale-infected animals maintained with ticks were positive 27 months after initial infection and a further 26 animals infected with A. marginale by blood inoculation were positive 3 months later. Three per cent of negative sera, 2% of B. bovis and 4% of B. bigemina-positive sera gave positive reactions.