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Related Experiment Videos

A sensitive solid phase enzymeimmunoassay for testosterone in plasma and saliva.

A Turkes, A O Turkes, B G Joyce

    Steroids
    |March 1, 1979
    PubMed
    Summary

    A new enzyme immunoassay (EIA) accurately measures testosterone in plasma and saliva. This sensitive method offers a reliable alternative to radioimmunoassay (RIA) for clinical diagnostics.

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    Area of Science:

    • Biochemistry
    • Endocrinology
    • Analytical Chemistry

    Background:

    • Accurate testosterone measurement is crucial for diagnosing various endocrine disorders.
    • Existing methods like radioimmunoassay (RIA) often require specialized facilities and handling of radioactive materials.
    • There is a need for sensitive, precise, and non-radioactive assays for testosterone determination.

    Purpose of the Study:

    • To develop and validate a sensitive solid-phase enzyme immunoassay (EIA) for determining testosterone concentrations.
    • To assess the suitability of the EIA for small plasma and saliva aliquots.
    • To compare the performance of the developed EIA with established methods like gas-liquid chromatography/mass spectrometry (GLC/MS) and RIA.

    Main Methods:

    • Development of a solid-phase antiserum using testosterone-11 alpha-hemisuccinate/bovine serum albumin conjugate coupled to cyanogen bromide activated cellulose.

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  • Preparation of an enzyme label using a covalently linked testosterone/horseradish peroxidase conjugate.
  • Validation of the assay for sensitivity, specificity, and precision using small plasma and saliva samples.

    • Main Results:

    • The developed EIA demonstrated high sensitivity with a lower limit of 4 pg/assay tube.
    • Assay results showed excellent agreement with both GLC/MS (r=0.96) and routine RIA (r=0.95).
    • Salivary testosterone concentrations in normal males correlated well with circulating levels, suggesting saliva as a viable alternative matrix.

    Conclusions:

    • The developed enzyme immunoassay is a sensitive, specific, and precise method for testosterone determination.
    • EIA offers a viable, non-radioactive alternative to RIA for both small clinical laboratories and high-throughput centers.
    • Saliva can be reliably used for testosterone level assessment in clinical studies, simplifying sample collection.