Development of a competitive immunoassay for the determination of cortisol in human saliva.

Adama Marie Sesay, Laura Micheli, Pirkko Tervo, Giuseppe Palleschi, Vesa Virtanen

CEMIS-Oulu Mesurement Technology Unit, Kajaani University Consortium, University of Oulu, 88600 Sotkamo, Finland.

Analytical biochemistry 2013 Mar 15

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Two new protein conjugates were prepared and studied to develop and compare two (direct and indirect) competitive enzyme-linked immunosorbent assay (ELISA) formats for the determination of cortisol in human saliva. Toward this goal, ovalbumin was conjugated to cortisol and used for developing an indirect competitive ELISA, while alkaline phosphatase was coupled with the same analyte for a direct competitive assay. The yield of the conjugation reactions was evaluated. The results obtained show that the indirect and direct ELISA formats developed for cortisol had working ranges of 0.5-70 and 2-330 ng/ml and detection limits of 0.5 and 1.2 ng/ml, respectively. Artificial and real saliva samples were spiked with cortisol to study the matrix effect of saliva. The suitability of the assays for quantification of cortisol in saliva was also studied. Copyright © 2012 Elsevier Inc. All rights reserved.

Citation

Adama Marie Sesay, Laura Micheli, Pirkko Tervo, Giuseppe Palleschi, Vesa Virtanen. Development of a competitive immunoassay for the determination of cortisol in human saliva. Analytical biochemistry. 2013 Mar 15;434(2):308-14