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Globally, the majority of new HIV infections are recorded in Eastern and Southern Africa, with the youth being disproportionately affected. HIV prevention is the cornerstone of controlling the spread of HIV and ending this epidemic by 2030. However, barriers to the utilization of HIV prevention services remained underexplored especially among the youth in rural settings in sub-Saharan Africa. This qualitative study, conducted between February and April 2022 in rural southwestern Uganda, explored these barriers and identified recommendations to improve the utilization of HIV prevention services among youth. We conducted six focus group discussions (with youth [15-24 years] both in and out of school), nine in-depth interviews (with teachers, health workers, and members of the village health team), and four key informant interviews (with district officials) to collect data. Thematic analysis revealed barriers at the individual level (e.g., misconceptions, fear of testing, low perceived HIV risk, confidentiality concerns), community level (e.g., stigma, lack of counseling, peer influence), and health system level (e.g., lack of youth-friendly services). Recommendations included formation of youth peer support groups, ongoing awareness campaigns, and socio-economic empowerment initiatives, particularly targeting adolescent girls and young women. National scaling of these initiatives is essential to overcoming identified barriers and reducing HIV transmission among this vulnerable population. Additionally, economic empowerment especially among adolescent girls and young women in rural areas has enormous potential to address the spread of HIV in this sub-population., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Ssali et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)