To detect ROS1 rearrangement in NSCLC, three different assays including fluorescent in stiu hybridization (FISH), Amoy Dx reverse transcription polymerase chain reaction (RT-PCR) and Oncomine Dx Target Test (ODXTT) were routinely performed as a nationwide screening In South Korea. ROS1 FISH or RT-PCR was consecutively and ODXTT was selectively performed in routine practice. The basic clinicopathologic and analytic characteristics were performed on whole cohort. 238 (1.8%) of 13,533 NSCLC confirmed positive for ROS1 rearrangement. Among above cohort, clinically selected 110 ROS1-negative cases confirmed by RT-PCR were re-evaluated by ODXTT and ODXTT found additional four ROS1-positive cases. one of two ROS1 positive cases by ODXTT confirmed responsive to crizotinib and the other showed no response to crizotinib. The average age of patients in the ROS1-positive group was 61 years old, Female patients (58.4%) with ROS1-positive occurred more than male patients (41.6%). Turnaround time for each method were 5.2 days by FISH, 4.8 days by RT-PCR, and three weeks by ODXTT, respectively.