To determine the prevalence and factors associated with the uptake of HIV self-testing (HIVST) among adolescent girls and young women (AGYW) attending middle learning institutions in Dodoma City, Tanzania, in 2024.

Design: A cross-sectional study design was employed.

The study was conducted in five randomly selected colleges in Dodoma City, Tanzania.

A total of 771 female students aged 15–24 years who provided informed consent were enrolled. Students who declined participation or were reported by the college matron as medically unfit were excluded. In this study, ‘sick’ referred to participants with a clinically diagnosed illness rendering them unable to participate.

The primary outcome was the uptake of HIVST among participants.

The mean age of participants was 20.78 years (SD=1.85). Overall, 360 participants (46.7% (95% CI 43.2% to 50.2%) reported having used HIVST. Among those who had not tested, the most commonly reported barriers included cultural resistance 392 (95.4%), fear of judgement from healthcare providers, 372 (90.5%); legal restrictions on kit provision, 360 (87.6%); fear of testing procedures, 291 (70.8%); concerns about test reliability, 286 (69.6%); fear of test results, 283 (68.9%); limited accessibility to HIVST kits, 280 (68.1%); fear of others discovering they had tested, 273 (66.4%); low awareness of HIVST, 193 (47.0%); and a perceived low risk of HIV infection, 73 (17.8%).

Factors significantly associated with HIVST uptake included having multiple sexual partners adjusted prevalence ratio (aPR 1.23, 95% CI 1.05 to 1.45), studying health-related courses (aPR 1.14, 95% CI 1.04 to 1.27), availability of kits (aPR 2.83, 95% CI 2.21 to 3.62), previous HIV testing experience (aPR 2.65, 95% CI 2.05 to 3.43) and perceiving oneself at risk of HIV infection (aPR 1.29, 95% CI 1.11 to 1.50).

The uptake of HIVST among AGYW in Dodoma City remains below the national target of 95% HIV awareness among people living with HIV. Uptake was influenced by factors such as multiple sexual partnerships, study discipline, kit availability, prior testing experience and perceived risk of infection. Addressing the identified barriers and improving awareness and accessibility of HIVST could enhance testing rates among AGYW.