One step enzyme linked immunosorbent assay for direct estimation of serum cortisol

Journal of Immunoassay
A Basu, Tulsidas G Shrivastav

Abstract

One step competitive enzyme linked immunosorbent assay (ELISA) for direct estimation of cortisol in human serum is described. Cortisol-3-O-carboxymethyl-oxime-bovine serum albumin (cortisol-3-O-CMO-BSA) was used as an immunogen and cortisol-21-hemisuccinate-horse radish peroxidase (cortisol-21-HS-HRP) was used as a tracer. To the cortisol antibody coated microtiter wells, standards or serum samples (25 microl) along with cortisol-HRP conjugate (100 microl) were incubated for 2 hours at 37 degrees C. Bound enzyme activity was measured by, using TMB/H2O2 as a substrate. In this new strategy, chilled acetone stripped pooled human serum and sodium salicylate were used for preparing the standards and blocking the cortisol binding globulin (CBG), respectively. The sensitivity of the assay was .28 microg/100ml. The intraassay and interassay coefficient of variations (CVs) were ranged from 1.3% to 9.3% and 6.8% to 12.3 %, respectively. The analytical recoveries were 94% to 101.5%. The serum cortisol values, obtained by this method were correlated well with those, obtained by radioimmunoassay; r=0.95 (n=52).

References

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Citations

Jun 5, 2003·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavKiran P Kariya
Jun 5, 2003·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavKiran P Kariya
Sep 5, 2003·Journal of Immunoassay & Immunochemistry·Tulsidas G Shrivastav
Oct 6, 2004·Journal of Immunoassay & Immunochemistry·Tulsidas G Shrivastav
Jul 7, 2011·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavRita Singh
Sep 30, 2008·Journal of Immunoassay & Immunochemistry·Seema NaraTulsidas G Shrivastav
Dec 13, 2005·Clinica Chimica Acta; International Journal of Clinical Chemistry·Anupam BasuTulsidas G Shrivastav
May 17, 2011·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavRita Singh
Nov 30, 2010·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavDinesh Kumar
Jun 29, 2012·Journal of Immunoassay & Immunochemistry·Tulsidas G ShrivastavDinesh Kumar

Related Concepts

Cortisol binding globulin
Antibody Specificity
Calibration
Carrier Proteins
Enzyme-Linked Immunosorbent Assay
Goat
Hydrocortisone, (9 beta,10 alpha,11 alpha)-Isomer
Radioimmunoassay
Regression Analysis
Intraobserver Variation

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