Objective To investigate the value of endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) cytopathological detection in diagnosis of lung cancer and sarcoidosis, and analyse the clinicopathological characteristics. Methods One hundred and sixty cases undergoing cytopathological detection were collected, and the value of cytopathological detection in the diagnosis of lung cancer and sarcoidosis was analysed based on the criteria of histologiopathology combined with clinical manifestations. Results The diagnosis of 150 cases was confirmed by histopathological examinations, including 108 cases of lung cancer, 23 cases of sarcoidosis, 4 cases of tuberculosis and 15 cases of the other diseases. Ninety-five cases of lung cancer were detected by EBUS-TBNA cytopathological detection, with the positive diagnostic rate of 88.0% (95/108) and negative diagnostic rate of 12.0% (13/108). The sensitivity, specificity, positive predictive value and negative predictive value of EBUS-TBNA cytopathological detection in diagnosis of lung cancer were 88.0%, 100%, 100% and 76.4% respectively. EBUS-TBNA cytopathological detection revealed 15 cases of non-caseous granuloma lesion among 23 cases of sarcoidosis, and 1 case of caseous granuloma in 4 cases of tuberculosis. Conclusion EBUS-TBNA cytopathology detection has high sensitivity and specificity in diagnosis of lung cancer, and may play a role in the diagnosis of sarcoidosis and tuberculosis.