This study evaluated the accuracy of NAVIFY Mutation Profiler, a cloud-based CE-IVD software that aids in interpreting clinically relevant variants detected in somatic oncology next-generation sequencing (NGS) tests. This tool reports tiered classifications based on different levels of clinical evidence from a highly curated, regularly updated database derived from medical guidelines, drug approvals, and peer reviewed literature. A retrospective analysis was performed on NGS results from 37 lung cancer cases treated with chemotherapy (n=10), EGFR tyrosine kinase inhibitor (TKI) (n=5), or ALK TKI (n=22). Several aspects were assessed, including accuracy of interpretation compared to manual curation, validity of curation content updates over time, and agreement with public databases. For chemotherapy cases with no targetable biomarkers, NAVIFY Mutation Profiler did not identify any targeted therapies. In EGFR and ALK TKI cases, the software associated appropriate targeted therapies and accurately interpreted variant combinations containing drug-resistance variants. Of the 9 unique ALK mutations conferring resistance to crizotinib, NAVIFY Mutation Profiler provided correct annotation for 9/9 mutations whereas OncoKB and COSMIC indicated crizotinib resistance for 8/9 mutations. Of all 145 variants analyzed, there was substantial agreement (Cohen’s kappa=0.62) between NAVIFY Mutation Profiler and OncoKB for classifying actionable mutations. Furthermore, NAVIFY Mutation Profiler presented accurate targeted therapies across different regions, e.g., European Union versus Canada, and remained up-to-date with evolving regional approvals and medical guidelines.Copyright © 2020. Published by Elsevier Inc.