Background: The aim of the study was to investigate the clinical utility of biomarkers, interferon gamma (INF-γ) and adenosine deaminase (ADA) in the differential diagnosis between tuberculosis and non-tuberculosis pleural effusion. Methods: The present study enrolled 130 patients with pleural effusion that were admitted to hospital between 2012-2015. From the patients were obtained pleural fluid and serum which were administrated to test analyses within 24 hours. Results: Based on Light's criteria, biochemical, cytological etc. analyses were established the pleural effusion as exudative and the etiology of the effusion, 40 malign, 48 tuberculosis, 42 parapneumonic. Pleural fluid, serum and pleural fluid/serum ADA and INF-γ pleural fluid results differ significantly between the different types of exudative effusion. For the difference of tuberculosis and non-tuberculosis pleural effusion ADA pleural fluid, ADA serum and INF-γ pleural fluid showed significance for this difference, meanwhile ADA pleural fluid/serum do not contribute in the difference. Conclusions: The study conducted that ADA and INF-γ markers can be used for the differential diagnosis of exudative pleural effusion and tuberculosis from non-tuberculosis. INF-γ even if it has a higher specificity than ADA, due to its cost we recommend that it cannot be promoted as the only solution that facilitate the different diagnosis, other markers need to be taken into consideration to assist the results.