The study of serum progesterone 17 hydroxyprogesterone and 17 beta estradiel by radioimmunoassay in Thai swamp buffalo

Abstract

A radioimmunoassay for the measurement of serum progesterone (P), 17 hydroxyprogesterone (17P) and 17B estradiol (E[subscript2]) were described. The antisera obtained from Dr. G.E. Abraham, Department of obstetrics and Gynecology, University of California. Male buffaloes free hormone serum (FHS) was prepared for pooled steroids preparation. Separation of free from bound hormone was achieved by dextran coated charcoal suspension. On ml of pooled serum with tritrated these steroids added for recovery, was extracted with ether, then chromatographed on a celite microcolumn. The specificity of antisera has been show significantly specific cross reaction. After purification step by celite chromatography, the contaminants could be all removed. Recovery of the labeled P, 17P and E[subscript2] after extraction and chromatography were 71.7–94.5%, 70.2–94.1% and 61.0–80.7% respectively. The percentage recovery of standard P, 17P and E[subscript2] added in FHS varied in the ranges of 72.9–89.6, 77.0–97.0 and 85.5-101.3 respectively. The precision of within and between assay variance was evaluated by duplicated measuraments of the same sample in the same assay and in 5 different assays. The coefficient of variation (CV) were 8.2% (P and 17P), 8.3% (E[subscript2]) and 13.2% (P), 11.2% (17P), 14.6% (E[subscript2]) respectively. The sensitivity varied between 3.5–25.0 pg of P, 4.0–25.0 pg of 17P and 2.5–10.0 pg of E[subscript2].