BACKGROUND: Gay, bisexual, and other men who have sex with men living with HIV (GBMSM-LWH) in the United States bear a heavy burden of bacterial sexually transmitted infections (STIs). Timely diagnosis and treatment are key to prevention. Only a few studies have combined home specimen self-collection for bacterial STI screening with live audio and video (AV) conferencing. None have focused on GBMSM-LWH or incorporated motivational interviewing (MI), a client-centered, strengths-based counseling approach that seeks to support individuals to create positive behavioral change. OBJECTIVE: Our study seeks to investigate the feasibility and acceptability of an MI-based telehealth intervention that integrates home specimen self-collection from different anatomical sites of possible exposure and MI delivered via live AV conferencing to engage sexually active GBMSM-LWH in bacterial STI screening. METHODS: Participants are being recruited from across the United States via advertising on mobile dating apps and social networking sites and via peer referral. Phase 1 involves piloting the delivery of an innovative telehealth intervention for bacterial STI screening to 75 GBMSM-LWH. Our intervention includes three components: (1) a pretest live AV conferencing session involving an MI-guided discussion to elicit awareness of bacterial STIs; fill any knowledge gaps; bolster the perceived importance of regularly testing for gonorrhea, chlamydia, and syphilis; and build self-efficacy for specimen self-collection; (2) home self-collection and return via mail of a urine sample (for gonorrhea and chlamydia testing), a throat swab (for gonorrhea and chlamydia testing), a rectal swab (for gonorrhea and chlamydia testing), and a finger-stick blood sample (for syphilis testing); and (3) a posttest live AV conferencing session involving an MI-guided discussion to prepare participants for receiving test results and formulate personalized action plans for seeking treatment (if warranted) and repeat testing. Descriptive statistics and progression ratios will be calculated, and potential variations in our intervention's feasibility and acceptability will be numerically summarized and graphically visualized. Phase 2 involves elucidating attitudes, facilitators, and barriers related to engaging in each intervention component via semistructured in-depth interviews with a purposive subsample of 20 participants who complete progressively smaller subsets of the pretest session, specimen return for bacterial STI testing, and the posttest session. Thematic analysis will be used to identify, analyze, and report patterns in the data. Quantitative and qualitative data will be integrated at the design, methods, interpretation, and reporting levels. RESULTS: Study procedures were approved by the Institutional Review Board at the University of Michigan in September 2023. Participant recruitment began in April 2024. CONCLUSIONS: Our study will advance multiple goals of the STI National Strategic Plan for the United States for 2021 to 2025, specifically those pertaining to preventing new STIs; accelerating progress in STI research, technology, and innovation; and reducing STI-related health disparities. TRIAL REGISTRATION: ClinicalTrials.gov NCT06100250; https://www.clinicaltrials.gov/study/NCT06100250. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/64433.