Development of a highly sensitive enzyme linked immunosorbent assay for human serum progesterone using penicillinase.

A simple, highly sensitive, heterologous, competitive, penicillinase based tube ELISA for human serum progesterone has been developed. Specific antisera against progesterone was raised in rabbits using 11 alpha - hydroxyprogesterone hemisuccinate-BSA. Progesterone-3-CMO was coupled to penicillinase by the carbodiimide method. The standard curve covered a wide range from 5 pg/tube to 800 pg/tube. The sensitivity of the assay was 3.8 pg/tube. The intrassay coefficient of variation ranged from 8.6% to 11.2% at low concentration while at medium and high concentrations it ranged from 5.3% to 8.9%. The interassay coefficient of variation for the low, medium and high concentrations of progesterone were 9.6%, 10.7% and 6.5% respectively. Parallelism between the standard curve and the sample dilution curve indicated accuracy of the assay. Samples measured by RIA and ELISA showed an excellent correlation (r = 0.98).