title: Development and validation of competitive ELISA to determine follicular fluid progesterone concentrations.
abstract: A simple, direct and reliable heterologous ELISA was developed and validated to determine follicular fluid progesterone concentrations in 10- to 20-mm antral follicles in heifers. A competitive ELISA was developed, using a policlonal antisera raised in New Zealand white rabbits and horseradish peroxidase as label. Standard curve covered a range between 0 and 1 ng per well. The intra- and inter-assay coefficients of variation (CV) were 6.54 and 8.27%, respectively (n = 10). The low detection limit was 1 pg per well with a sensitivity of 50% binding 83.17 pg per well. Comparison of ELISA with RIA showed a correlation coefficient of 0.98. A total of 34 follicles obtained from heifers in the follicular phase of the estrous cycle were tested, and the mean progesterone concentration was 86.72 +/- 20.39 ng/ml. These results were similar to those previously reported for the same species, age, follicular size and stage of cycle.
authors.affiliation: Departamento de Fisiología Animal Facultad de Veterinaria, UCM 28040 Madrid, Spain.
authors.forename: G
authors.initials: G
authors.lastname: Silván
authors.forename: J C
authors.initials: JC
authors.lastname: Illera
authors.forename: M
authors.initials: M
authors.lastname: Illera