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Objective: The aim of this study was to evaluate the reliability of a non-invasive method (saliva) for determination of testosterone level in diagnosing cases of Polycystic Ovarian Syndrome (PCOS), and comparing it with serum free testosterone. Study design: Prospective clinical controlled trial. Participants and interventions: Fifty women with PCOS and 20 normal women as control group were selected for the study. They all aged between 23 and 35year. For all patients serum follicle stimulating hormone (FSH) and luteinizing hormone (LH) were assessed by chemiluminescent assay. Serum free testosterone (FT) was assayed by radioimmunoassay (RIA) and salivary testosterone (ST) by ELISA technique. BMI was calculated for all participants and transvaginal sonography to determine ovarian morphology and ovarian volume. Results: The present study showed a significant positive correlation between salivary testosterone, as measured by ELISA, and serum free testosterone measured by RIA (p=0.001 and r=0.52). Using the receiver operator curve, salivary testosterone was found to be more sensitive than serum (FT) (84% vs. 66%) in diagnosing PCOS patients. Conclusion: Determination of salivary testosterone is a reliable method to detect changes in the concentration of available biologically active testosterone in the serum. Salivary testosterone provides a sensitive, simple, reliable, non-invasive and uncomplicated diagnostic approach for PCOS.