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Related Experiment Video

Updated: Jul 17, 2026

Immunoblot Analysis
16:01

Immunoblot Analysis

Published on: June 20, 2008

Enzyme immunoassay of testosterone using nitrocellulose discs as the solid phase.

A K Samanta1, E Ali

  • 1Indian Institute of Chemical Biology, Calcutta.

Journal of Clinical Chemistry and Clinical Biochemistry. Zeitschrift Fur Klinische Chemie Und Klinische Biochemie
|December 1, 1990
PubMed
Summary

A new enzyme immunoassay (EIA) offers a cost-effective method for measuring serum testosterone. This paper disc-based assay provides accurate results comparable to radioimmunoassay (RIA), with convenient sample handling.

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Area of Science:

  • Biochemistry
  • Immunology
  • Analytical Chemistry

Background:

  • Accurate measurement of serum testosterone is crucial for diagnosing various endocrine disorders.
  • Existing methods like radioimmunoassay (RIA) can be expensive and time-consuming.
  • Development of a simpler, more cost-effective immunoassay is desirable.

Purpose of the Study:

  • To develop and validate a novel enzyme immunoassay (EIA) for serum testosterone measurement.
  • To utilize nitrocellulose paper discs as a solid support for the EIA.
  • To evaluate the assay's performance against established methods.

Main Methods:

  • Developed an enzyme immunoassay (EIA) using testosterone-specific antibody-coated nitrocellulose paper discs.
  • Incubated discs with serum samples, testosterone, and a testosterone-peroxidase conjugate.

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  • Quantified bound peroxidase activity to determine testosterone levels.

    • Main Results:

    • The EIA showed good correlation with radioimmunoassay (RIA) (correlation coefficient = 0.84).
    • The assay demonstrated minimal cross-reactivity with other steroids, except dihydrotestosterone.
    • Reported inter- and intra-assay coefficients of variation of 4.4% and 9.6%, respectively.

    Conclusions:

    • The developed paper disc-based EIA is a reliable and cost-effective method for serum testosterone measurement.
    • The assay offers convenient preservation and transport of reagents.
    • This method presents a viable alternative to traditional RIA and ELISA for testosterone quantification.