OBJECTIVES:  Evaluate the budget impact of next-generation sequencing (NGS) instead of single-gene testing for tissue-based molecular assessment of non-squamous, advanced non-small cell lung cancer (aNSCLC) from the United States healthcare payer perspective. METHODS:  A Markov model was developed to evaluate an annual cohort of newly-diagnosed, non-squamous aNSCLC patients in a hypothetical million-member plan followed for 5 years. aNSCLC epidemiology data were from published literature. Mutation prevalence and testing rates for activating mutations were considered for current and emerging gene targets (EGFR, ALK, ROS-1, BRAF, MET, HER2, RET), and sourced from literature. Rates of successful test completion were informed by the NCI-MATCH trial and experience at a large commercial laboratory. Patients identified as EGFR+ and ALK+ were assumed to be treated with erlotinib and crizotinib, respectively; clinical trial enrollment was an option for patients with other mutations. Adverse events, progression, and survival rates with targeted therapy or chemotherapy were from randomized clinical trials. Costs of testing and first-line and maintenance therapies were based on Medicare 2016 reimbursement values; adverse event and post-progression costs were from literature. RESULTS:  Of 1-million plan members, 318 patients were expected to be newly-diagnosed with non-squamous aNSCLC and 180 were tested. Of the 58 tested patients expected to have activating mutations, single-gene testing identified 35 patients and NGS identified 54. Testing-related costs decreased $47,870 with NGS instead of single-gene testing. First-line and maintenance treatment costs increased $841,894, offset by $391,891 of savings in post-progression-related costs. Total budget impact over 5 years for this cohort assessed with NGS instead of single-gene testing was $402,133 ($0.0067 per-member per-month). CONCLUSIONS:  NGS is expected to improve identification of activating mutations and enable improved patient selection for targeted therapy and/or clinical trial enrollment. The impact to payer costs is expected to be minimally cost-additive.