Competitive chemiluminescent enzyme immunoassay for testosterone

Objective: To establish a chemiluminescent enzyme immunoassay (CLEIA) for testosterone. Methods: Antibody was coated and antigen was labeled or antigen was coated and antibody was labeled. Horseradish peroxidase was used as a label and enhanced chemiluminescent system as substrate. Results: The method with coated second antibody and labeled testosterone had high sensitivity and simple measures. The sensitivity of the assay was 0.063 ng/ml. The testosterone was added to the steroidfree serum at concentrations of 1.0, 5.0 and 10.0 ng/ml, the withinrun coefficients of variation were 5. 8%7.4%( n =20), and runtoruns' were 6.6%8.7%( n =2)respectively. The recovery rates were from 98.2% to 104.0%. The assay was not crossreacted significantly with other steroids except 5dihydrotestosterone with 20% cross reactivity. It correlated well with Boehringer Mannheim enzyme immunoassay kit (BM) for testosterone. CLEIA=0.915 BM+0.598 ng/ml, n=128, r=0.971. Conclusion: The chemiluminescent enzyme immunoassay has high sensitivity, good accuracy, repeatability and correlation with commercial kit. The method is helpful to clinical laboratory diagnosis.