Application of the enzyme-linked immunosorbent assay to detect canine erythrocyte antibodies.

A double-antibody enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies bound to canine RBC. The method used a horseradish peroxidase conjugate, o-dianisidine, as a water-soluble chromogen and osmotically lysed RBC on which antibody was detected. The ELISA method was compared with the traditional antiglobulin (Coombs') test on positive controls, using sheep RBC sensitized with dilutions of canine anti-sheep RBC serum and on RBC from canine patients tested for autoantibodies to RBC. The ELISA method appeared to be more sensitive than the Coombs' test and gave an estimate of the amount of antibody present without the need for serial antiserum dilutions to determine the antibody titer.