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A simple direct solid-phase enzymeimmunoassay for cortisol in plasma.

R K Hindawi, S J Gaskell, G F Read

    Annals of Clinical Biochemistry
    |January 1, 1980
    PubMed
    Summary
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    A new enzyme-linked immunoassay for plasma cortisol offers a simple and accurate method for its determination. This assay is suitable for routine clinical use, especially in labs with variable workloads.

    Area of Science:

    • Clinical Chemistry
    • Biochemistry
    • Immunoassay Development

    Background:

    • Plasma cortisol measurement is crucial for diagnosing adrenal disorders.
    • Existing methods like radioimmunoassay and gas chromatography-mass spectrometry can be complex or require specialized equipment.
    • A need exists for a simpler, more accessible assay for routine clinical use.

    Purpose of the Study:

    • To develop and validate a direct solid-phase enzyme-labelled immunoassay for plasma cortisol.
    • To assess the assay's sensitivity, accuracy, and precision.
    • To evaluate its applicability in routine clinical settings.

    Main Methods:

    • A solid-phase enzyme immunoassay was established using horseradish peroxidase/cortisol conjugate as the enzyme label.
    • Antiserum was coupled to cellulose for efficient separation of free and bound steroids.

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  • Heat denaturation was employed to eliminate the need for solvent extraction of cortisol-binding globulin.

    • Main Results:

    • The developed enzyme immunoassay demonstrated a lower limit of sensitivity of 16.6 nmol/l.
    • The assay met standard criteria for accuracy and precision.
    • Results showed excellent agreement with gas liquid chromatography/mass spectrometry (r=0.98) and radioimmunoassay (r=0.95).

    Conclusions:

    • The developed enzyme immunoassay provides a simple, direct, and accurate method for plasma cortisol determination.
    • The assay is particularly suitable for routine analysis in small clinical laboratories or those with fluctuating workloads.
    • This method offers a practical alternative to more complex diagnostic techniques.