Development of a Semi Quantitative Progesterone Enzyme Immunoassay for Determination of Ovulation Time in Bitches

A solid phase residual binding progesterone enzyme immunoassay (EIA) for determination of ovulation time in bitches was developed using polyclonal antibodies against progesterone-BSA and progesterone-3-CMO-HRPO conjugate. The assay was optimized with 1:100 dilution of progesterone polyclonal antibody, 1:1000 dilution of progesterone -3-CMO HRPO conjugate and 1:10 dilution of serum samples. Validation of the developed assay for its sensitivity, specificity and accuracy by comparing it with vaginoscopy showed an overall agreement of 97 per cent between EIA and vaginoscopy. The diagnostic sensitivity of the progesterone EIA in predicting the preovulatory, ovulatory and post ovulatory periods were 96.42, 91.66 and 100 per cent respectively. The diagnostic specificity of the assay was 100 per cent. Comparison of the developed EIA with Chemiluminescence assay (CIA) gave a correlation coefficient (r) of 0.92.