BACKGROUND: HIV remains a significant global public health challenge, disproportionately affecting adolescent girls and young women (AGYW). Uganda has adopted different strategies of HIV Testing Services (HTS) to improve access and utilization, but the uptake of these services by AGYW is still low. This study explored barriers, facilitators, and preferences for HTS among AGYW in Rakai district, Central, Uganda. METHODS: This qualitative study employed in-depth interviews with 24 purposively selected AGYW aged 15-24 years who had used HTS at least once in the past year. Participants were drawn from diverse backgrounds based on age, residence (fishing or mainland communities), schooling, employment, and marital status. Data were analyzed thematically using both inductive and deductive approaches in Atlas.ti software. RESULTS: Key barriers to HTS uptake varied by age and residency. Adolescent girls aged 15-19 years, especially those in school, expressed strong fear of blood-based testing and discomfort with invasive procedures, while young women aged 20-24 years emphasized long waiting times, provider attitudes, and confidentiality concerns, particularly in public facilities. AGYW from fishing communities reported heightened stigma and privacy challenges in public facilities, making them more reliant on discreet mobile and outreach services. In contrast, mainland residents favored private facilities for their efficiency. Facilitators across all groups included mobile HTS and community outreaches, which improved accessibility and reduced logistical barriers to utilization. Younger adolescents preferred saliva-based, less invasive tests, while older participants favored facility-based testing for the professional support it offered. Preferences also varied across provider characteristics: married and older AGYW favored experienced adult providers, while younger participants, particularly in mainland areas, expressed comfort with male providers seen as more empathetic. CONCLUSION: HTS strategies should offer a diverse range of service delivery options that reflect the varied preferences and needs of AGYW by age and residency. Expanding mobile testing, offering less invasive options, ensuring confidentiality, and improving provider interactions are critical for increasing HTS uptake in high-risk settings like Rakai.