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Objective: To determine whether endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) can rapidly distinguish among lung cancer, tuberculosis and sarcodosis, and to explore its sensitivity and specificity. Methods: Clinical data of patients with enlarged mediastinal or hilar lymphadenopathy who underwent EBUS-TBNA at our hospital between September 1, 2012 and June 30, 2015 for were retrospectively analyzed. The sensitivity, specificity, positive predictive value, and negative predicted value [including 95% confidence interval (CI)] were calculated.. Results: A total of 299 lymph nodes from 201 patients underwent EBUS-TBNA were selected and no serious complications occurred. EBUS-TBNA showed a sensitivity of 87.9% (124/141) (95% CI: 81%–92%), a specificity of 100.0% (124/124) (95% CI: 97%–100%), and a negative predicted value of 41.3% (95% CI: 23%–61%) in the detection of lung cancer. The sensitivity and specificity of diagnosis of mediastinal tuberculosis lymphadenitis were 72.4% (21/29) (95% CI: 53%–87%) and 100.0% (95% CI: 82%–100%); while sensitivity and specificity of diagnosis of sarcoidosis were 71.4% (5/7) (95% CI: 29%–96%) and 100.0% (95% CI: 91%–100%). Conclusions: The sensitivity and specificity of EBUS-TBNA do not significantly differ for a diagnosis of lung cancer versus tuberculosis or sarcodosis. [ABSTRACT FROM AUTHOR]