BackgroundAdolescents and young adults (AYA) in the United States, and in Washington, District of Columbia (DC), specifically, are disproportionately affected by HIV. Both the national Ending the HIV Epidemic initiative and DC-specific plans emphasize HIV testing, and innovative strategies to encourage testing among AYA are needed.ObjectiveThe purpose of this study is to identify sexual behaviors, HIV knowledge, HIV perceptions (eg, susceptibility and severity), and perceived barriers and facilitators to HIV testing among AYA at risk for HIV in DC.MethodsThis study was part of a larger study to determine the acceptability of using a life-and-dating simulation game to increase HIV testing among AYA. Focus groups and surveys stratified by self-reported sexual orientation were conducted among, and administered to, AYA aged 13-24 years in DC. HIV knowledge was explored during focus groups and measured using an adapted version of the Brief HIV Knowledge Questionnaire. Survey data were summarized using descriptive statistics and compared by self-reported sexual orientation. Transcripts were thematically analyzed.ResultsOf the 46 AYA who participated in the focus groups, 30 (65%) identified as heterosexual and 16 (35%) as lesbian, gay, bisexual, transgender, or queer. A higher proportion of lesbian, gay, bisexual, transgender, or queer AYA reported sexual activity (12/16, 75%, vs 18/30, 60%), condomless sex (11/12, 92%, vs 15/18, 83%), and HIV testing (13/16, 81%, vs 17/29, 58%) than heterosexual AYA. HIV prevention (“condoms” and “...PrEP”) and transmission (“exchange of fluids”) knowledge was high, and most (34/44, 77%) of the AYA perceived HIV testing as beneficial. However, the AYA also demonstrated some misinformation concerning HIV: an average of 67% (31/46; SD 0.474) of the participants believed that an HIV test could deliver accurate results 1 week after a potential exposure and an average of 72% (33/46; SD 0.455) believed that an HIV vaccine exists. The AYA also identified individual (“...people...are scared”), interpersonal (“it’s an awkward conversation”), and structural (“...people don’t...know where they can go”) barriers to testing. Most of the AYA indicated that they were very likely to use the demonstrated game prototype to help with getting tested for HIV (median 3.0, IQR 2.0-3.0, using a scale ranging from 0 to 3, with 3 indicating high likelihood) and strongly agreed that the game was interesting (median 5.0, IQR 5.0-5.0), fun (median 5.0, IQR 4.0-5.0), and easy to learn (median 5.0, IQR 5.0-5.0, using a scale ranging from 1 to 5, with 5 indicating strong agreement).ConclusionsThese results suggest a need for multilevel HIV testing interventions and informed the development of a mobile health intervention aiming to increase HIV knowledge and risk perception among AYA, while reducing barriers to testing at the individual and structural levels, supporting efforts to end the domestic HIV epidemic.