A Sensitive Direct Enzyme Immunoassay for Cortisol in Plasma and Saliva1)

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Abstract
A sensitive enzyme immunoassay for cortisol in plasma and saliva without extraction was developed. An antibody was raised in rabbits against cortisol-3-CMO-BSA and compared with anti-cortisol-21-HS-HSA. The enzyme label was a covalently linked cortisol-horseradish peroxidase conjugate. Solvent extraction was avoided by use of heat denaturation of transcortin and dilution of plasma 1:300. This assay had a limit of detection of 3 fmol/tube and satisfied the standard criteria of accuracy and precision. The test requires only a simple photometer. The normal values of cortisol in plasma and saliva before and after ACTH, dexamethasone or metyrapone application were determined. Saliva, being easily collected by a noninvasive, stress-free technique is an alternative to plasma. The cortisol concentrations were significantly increased in plasma and saliva of patients with Cushing''s syndrome and decreased in patients with Addison''s disease.