Abstract: To evaluate the role of real-time endobronchial ultrasound-guided transbronchial needle aspiration ( EBUS-TBNA ) in the diagnosis of lung cancer. Methods: A total of 46 patients with mediastinal, and/or hilar lymphadenopathy, and/or intrathoracic peritracheal masses ( ≥1 cm ) previously detected by CT or PET-CT scan and who underwent EBUS-TBNA between August 2010 and February 2011 were retrospectively reviewed. Twenty-five cases were diagnosed as lung cancer with hilar and/or mediastinal lymph node metastasis, and 21 cases were unknown mediastinal and/or hilar lymphadenopathy. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnosis rate of EBUS-TBNA in the diagnosis of lung cancer were calculated. Results: Among the 46 patients, pathological examination diagnosed 38 cases of lung cancer, 3 of lymphatic tuberculosis, 3 of lymphadenitis, 1 of sarcoidosis, and 1 of lymphoma. EBUS-TBNA diagnosed 34 cases of lung cancer, 2 of lymphatic tuberculosis, 3 of lymphadenitis, and 1 of sarcoidosis. In 38 lung cancer cases, 48 samples were obtained from lymph nodes, and 1 sample was obtained from an intrapulmonary lesion. A total of 34 cases were diagnosed by EBUS-TBNA, and false negatives were obtained in 4 cases. The sensitivity, specificity, positive predictive value, negative predictive value, and diagnosis rate were 89.5%, 100%, 100%, 66.7%, and 87%, respectively. The procedure was safe and no severe complication occurred. Conclusion: The diagnosis rate, sensitivity, and negative predictive value of EBUS-TBNA are high. Therefore, EBUS-TBNA may be a safe and effective technique in the diagnosis of lung cancer.