An enzyme immunoassay for the quantitative determination of cortisol in serum and plasma.

Cortisol/hydrocortisone, compound F) is the main corticosteroid secreted in humans by the adrenal cortex. This steroid hormone has a molecular weight of 363.5. In most physiological conditions, only about 10% of plasma cortisol circulates unbound from transcortin and albumin. Among the products of the human adrenal cortex, only cortisol is involved in the regulation of ACTH secretion. As the level of free (non-protein bound) cortisol in blood rises, the release of ACTH is inhibited by the negative feedback effect. Conversely, if cortisol levels are subnormal, the negative feedback decreases, ACTH levels rise, and the adrenal cortex secretes cortisol until normal blood levels are restored. The release of ACTH is under control of hypothalamic corticotrophin-releasing hormone (CRH); the negative feedback system involving cortisol has been identified at both hypothalamic and pituitary levels. Normally during the day there is a fluctuation of cortisol achieving the highest level in the morning and the lowest in the night. Useful information is given when cortisol measurement is done in samples withdrawn at a fixed hour (8.00 a.m.). The main biological effects of cortisol are:promotion of gluconeogenesis, deposition of liver glycogen, increase in blood glucose concentration when the carbohydrate utilization is reduced, effect on fat metabolism and anti-inflammatory action. Cortisol measurement is a powerful tool for the evaluation of suspected abnormalities in glucocorticoid production: CushingÕs Syndrome (hypercortisolism), AddisonÕs disease or secondary adrenal insufficiency (hypocortisolism). In many cases, it is necessary to perform dynamic tests (suppression or stimulation) in order to localize the defect at one of the three main levels (i.e. adrenal, pituitary, hypothalamus).

The DRG¨ Cortisol ELISA Kit is a solid phase enzyme-linked immunosorbent assay (ELISA), based on the principle of competitive binding. The microtiter wells are coated with a monoclonal antibody directed towards an antigenic site on the Cortisol molecule. Endogenous Cortisol of a sample competes with a Cortisol-horseradish peroxidase conjugate for binding to the coated antibody. After incubation the unbound conjugate is washed off.The amount of bound peroxidase conjugate is inversely proportional to the concentration of Cortisol in the sample. After addition of the substrate solution, the intensity of color developed is inversely proportional to the concentration of Cortisol in the sample.