Abstract: Objective To evaluate the diagnostic value and safety of non-real-time endobronchial ultrasound （EBUS） guided transbronchial lung biopsy （TBLB） for the diagnosis of peripheral lung cancer. MethodsFrom December 2014 to June 2016，142 patients without obvious pathological changes in ophthalmic examinations by routine bronchoscopy were diagnosed as peripheral lung cancer in Fuzhou Pulmonary Hospital by EBUS， and their clinical data were retrospectively analyzed. In this study，102 cases of peripheral lung cancer patients received non-real-time EBUS guided TBLB，and 42 patients received routine TBLB. The diagnostic positive rate， lesion detection and safety were compared between non-real-time EBUS guided TBLB and conventional TBLB. Moreover，the factors that affect diagnosis positive rate were summarized. Results Among 142 patients receiving EBUS examinations， 71.8%（102/142） of pulmonary lesions were detected by EBUS. The dignostic rate of the EBUS guided TBLB was 69.6% （71/102）， higher than 15.0% （6/40） of conventional TBLB （P<0.05）. The dignostic rate of EBUS guided TBLB on lesions with diameter ≤ 20 mm was 50.0% （11/22），lower than 75.0% （60/80） on lesions with diameter >20 mm （P<0.05）. The dignostic rate of EBUS guided TBLB on probe completely envelop wind the lesions was 84.4% （38/45），higher than 57.9% （33/57） on probe incompletely envelop wind the lesions （P<0.05）. The dignostic rate of EBUS guided TBLB on lesions with the presence of a bronchus sign on HRCT imaging was 82.4% （28/34）， higher than 63.2% （43/68） on lesions without the presence of a bronchus sign on HRCT imaging（P<0.05）. Complications of biopsy occurred in this study included slight haemoptysis and chest pain without pneumothorax. The incidence rates of haemoptysis were 30.3% （43/142） in EBUS guided TBLB and 32.5% （13/40） in conventional TBLB （P>0.05）. The incidence rates of chest pain were 12.7% （18/142） in EBUS guided TBLB and 10.0%（4/40） in conventional TBLB （P>0.05）. Patients with these complications recovered spontaneously without special managements. Conclusion The procedure of non-real-time EBUS guided TBLB was minimally invasive and had higher diagnostic rate with fewer complications. It was a safe and effective method to diagnose peripheral lung cancer，while careful selection of suitable case could further improve the diagnostic accuracy.