Background: Although tuberculosis is a known disease, still on occasion, the tuberculosis patients remain undiagnosed and hence not treated appropriately. In some cases, the patient is not able to give a proper sputum sample. In some other cases, expert laboratory technician to examine the patient's sputum smear is not available. Furthermore, and in most cases of extrapulmonary tuberculosis, invasive diagnostic procedures such as biopsy might be followed. The aim of this study was to investigate the diagnostic value of serum level of adenosine deaminase (ADA) in extra pulmonary tuberculosis. Methods: In this analytical study, pulmonary tuberculosis patients were compared with two other groups of patients including lung cancer patients and healthy controls, based on national tuberculosis protocol. In each group, 40 patients who almost matched for age and sex were included. Blood samples were taken from the participants to measure serum ADA levels. Data were analyzed using SPSS software version 15.0. Results: Based on the data analyzed in this study, the mean ADA level in the patients with extra pulmonary tuberculosis was 23.8 IU/L which was significantly higher than that in the other groups (P < 0.001). Mean ADA levels in lung cancer patients and in healthy subjects were 15.8 IU/L and 10.7 IU/L, respectively. Conclusions: According to the results of this study, higher cutoff value for ADA would increase the specificity up to 100%. Moreover, serum ADA level can be a valuable additional index in diagnosis of extra pulmonary tuberculosis.