ImportanceAmid rapid and widespread adoption of telehealth-based opioid treatment (TBOT), there is an urgent need for rigorous studies exploring the feasibility and characteristics of urine drug screening (UDS).ObjectiveTo investigate administration patterns and results of UDS to assess feasibility of UDS and patient outcomes in a TBOT setting.DesignThis observational cohort study was conducted between January 1, 2021, and December 6, 2022, and included patients with opioid use disorder treated in Ophelia, a TBOT treatment platform in 14 states. Data analysis was performed from January to March 2023.Main Outcomes and MeasuresNumber and percentage of patients with UDS within 30, 90, and 180 days of intake, grouped by adherence to clinical protocols. Associations were assessed between baseline characteristics and UDS completion and opioid positivity in first 30 days using χ2 tests. Baseline and 180-day follow-up UDS results were compared using McNemar tests.ResultsAmong 3395 patients (mean [SD] age, 38.2 [9.3] years, mostly male [54.1%], non-Hispanic White [81.5%], urban-residing [80.3%], and cash-pay at intake [74.0%]), 2782 (83.3%) completed a UDS within 30 days (90.0% among protocol-adherent patients, 67.0% among protocol-nonadherent patients). A total of 2750 of 2817 (97.6%) patients retained more than 90 days completed 1 or more UDS, as did 2307 of 2314 (99.7%) patients retained more than 180 days. Younger patients, patients of a racial and ethnic minority group, those living in urban areas, and cash-pay patients were less likely to complete a UDS in the first 30 days. Buprenorphine positivity increased (from 96.9% to 98.4%, P = .004) and opioid positivity declined (from 7.9% to 3.3%, P < .001) over time.Conclusions and RelevanceIn this cohort study of patients with opioid use disorder receiving buprenorphine in a remote care environment, UDS was highly feasible, though early UDS completion rates varied across demographic subgroups. The prevalence of unexpected UDS results was low and declined over time in treatment.