Introduction The integration of telehealth into clinical care has increased access to substance use disorder (SUD) treatment. However, drug screen monitoring has largely remained tied to in-person visits, preventing a fully virtual workflow. We evaluated the acceptability, appropriateness, feasibility, and usability of video-observed oral fluid (VOOF) self-collection as an approach to address this implementation gap. Methods Participants receiving SUD treatment in outpatient ( n = 13) or residential ( n = 15) settings completed a video-recorded oral fluid self-collection protocol, followed by the Acceptability of Intervention Measure, Intervention Appropriateness Measure, Feasibility of Intervention Measure, and System Usability Scale. Results Most participants had multiple SUDs ( n = 23, 82.1%), and nearly all had co-occurring mental illness ( n = 27, 96.4%). The majority completed study measures ( n = 23, 82.1%) and returned collection kits ( n = 25, 89.2%). Acceptability, appropriateness, and feasibility all exceeded established thresholds (>3), with respective means of 3.5 (SD 1.1), 3.8 (SD 1.1), and 3.4 (SD 1.3). Usability scores were below average and more variable (mean = 51.8, SD 20.4). Scores did not significantly differ by care setting. Discussion This early-stage pilot found that VOOF self-collection was generally acceptable, appropriate, and feasible among individuals with SUD in real-world care settings. Usability scores were below average, with technical challenges related to video recording and upload likely contributing. Integrating VOOF within digital treatment platforms may streamline the user experience and improve usability while better aligning drug screen monitoring with the growth of telehealth in SUD treatment. This approach may benefit individuals experiencing transportation barriers and those in rural areas who live significant distances from testing sites.