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

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.
• 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.