Direct ELISA for estrone measurement in the feces of sows: prospects for rapid, sow-side pregnancy diagnosis.
Both free and conjugated fecal estrogens were surveyed in sows by means of RIA's after extraction and column chromatography. Six different RIA's were performed using the same fecal suspension. Based on differences in concentration in feces from 6 pregnant and 4 nonpregnant sows, estrone (E1) was selected for the development of a homologous, competitive ELISA for pregnancy diagnosis purposes. Polyclonal antibodies were raised against 6-keto-E1-carboxymethyloxime coupled to BSA. Biotinylated E1 was used as the competitive agent and was detected with a streptavidin-peroxidase conjugate. Tetramethylbenzidine was used as chromogen. Validation of this direct, easy to perform ELISA showed satisfactory specificity, accuracy, precision and sensitivity. Comparison of the assay with a validated RIA (including extraction and column chromatography) resulted in a linear regression equation of ELISA = 0.88 RIA - 0.72 (r = 0.95; n = 24 fecal samples). The E1 values in feces from 11 pregnant sows during the first 40 d of gestation compared with values from 10 nonpregnant sows showed significant differences between Days 23 and 35 of pregnancy (P < 0.05). The highly significant difference in E1 concentrations (P < 0.0012) between Days 27 and 30 in particular offers promise for pregnancy diagnosis.
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DLO-Institute for Animal Science and Health, Department of Reproduction, P.O. Box 65, NL 8200 AB Lelystad, The Netherlands.