OBJECTIVE: Assess the diagnostic yield of real-time bronchoscopic ultrasound transbronchial needle aspiration (EBUS TBNA) in conventional TBNA-negative mediastinal lymphadenopathy and hilar lymphadenopathy. MATERIALS AND METHODS: Sixty-two patients having either conventional TBNA-negative mediastinal lymphadenopathy or hilar lymphadenopathy underwent real-time EBUS TBNA. RESULTS: EBUS TBNA was performed on 72 lymph nodes (mediastinal = 48; and hilar = 24). 31 of the 72 (43%) lymph node samples were positive for malignancy (29) or benign diagnosis (2), and 17 of the 72 (24%) lymph nodes were true negative at EBUS TBNA confirmed at surgery. Out of 48 mediastinal lymph nodes EBUS TBNA was diagnostic for malignancy in 19 (40%) and negative in 14 of which 12 (86%) were surgically confirmed true negative and 2 (14%) false negative. In 10 of the 24 (42%) hilar lymph nodes, EBUS TBNA was diagnostic for malignancy while 5 were true negative and 1 false negative. All false-negative lymph nodes were PET positive. Adequacy of EBUS TBNA based on positive aspiration and surgically confirmed true negative was 67% and in patients suspected for malignancy was 77%. CONCLUSIONS: EBUS TBNA has a good diagnostic yield in hilar lymphadenopathy and in conventional TBNA-negative mediastinal lymphadenopathy.