Objective To evaluate the diagnosis value of radial probe endobronchial ultrasound guide sheath transbronchial lung biopsy (RP-EBUS-GS-TBLB) combination with rapid on-site evaluation (ROSE) in peripheral pulmonary lesions (PPLs). Methods One hundred and fifty-eight patients with PPLs identified by computed tomography in Nanjing Chest Hospital underwent RP-EBUS-GS-TBLB with or without ROSE randomly between February 2016 and August 2017. The sensitivity, the procedure time, the biopsy times, and the complications were evaluated in the two groups. Results The diagnostic yield was 85.7% (72/84) in ROSE group and 70.3% (52/74) in No-ROSE group. There was significant difference in diagnostic sensitivity between the two groups (P<0.05). The mean procedure time and number of biopsy in ROSE group were less than those in No-ROSE group (P<0.01). No severe procedure related complications such as pneumothorax and hemoptysis were observed. Conclusions ROSE can improve the diagnostic sensitivity, and shorten the procedure time. RP-EBUS-GS-TBLB combined with ROSE is a safe and effective technique for PPLs.