Comparison of an automated radioimmunoassay method and a competitive protein-binding method for evaluation of serum...

A competitive protein-binding method (CPB) for serum cortisol determination has been compared with a newly developed automated radioimmunoassay (RIA) technic with respect to reproducibility and correlation of patient results. The RIA method uses an immobilized anti-cortisol antibody covalently coupled to the inner surface of a plastic cartridge. A mixture of 3H-labeled and unlabeled cortisol is passed through the cartridge to achieve separation of free and bound cortisol. The bound cortisol is later eluted to regenerate the antibody for the next sample. The RIA procedure had a day-to-day coefficient of variation (CV) of 7% at three cortisol levels. The CPB method had a day-to-day CV of 10% at a concentration of 8 microgram/dl. A comparison of 67 patient samples showed no statistically significant difference between the methods. The regression equation was RIA = 0.96 CPB + 0.14 microgram/dl, with a correlation of 0.92.