Background The Human Papillomavirus (HPV) vaccine has offered a great promise to reduce the cervical cancer burden; its utilization (uptake) however has been lagging. However, the levels and factors associated with the uptake of the vaccine have not been well investigated, especially in the local context.

Objective To assess the uptake of human papillomavirus vaccination and its associated factors among adolescent school girls in ambo town, Oromia, Ethiopia, 2020.

Methods An institution-based cross-sectional quantitative study design supplemented with the qualitative inquiry was employed to assess Human Papillomavirus vaccination uptake and its associated factors among 422 adolescent school girls in Ambo town, central Ethiopia from December 1–30, 2020. The collected data were coded, entered, and cleaned by using Epi info 7.2.3 and exported to SPSS version 25 for analysis. Descriptive statistics were used to compute summary statistics and proportions. Both bivariate and multivariable logistic regression was employed to identify factors associated with HPV vaccine uptake. Adjusted odds ratio and 95% confidence interval were used for the strength and directions of association. A P-value of < 0.05 was used to declare statistical significance. Qualitative findings have been analyzed with manual thematic analysis.

Result The proportion of HPV vaccination uptake among school girls in this study was 44.4%. Hearing about HPV vaccine [AOR = 2.50, 95%CI: (1.045–5.959)], availability of awareness creation [AOR = 2.53, 95%CI: (1.507–4.258)], and favorable attitude [AOR = 2.049, 95%CI: (1.153–3.64)] were the key identified factors associated with vaccination uptake. In addition, poor perception, fear of side effects, and misunderstanding were among the major factors identified by qualitative findings.

Conclusion There was low uptake of HPV vaccination among the school Adolescents in the study area. Availability of awareness creation programs, favorable attitude towards HPV vaccine, and hearing about HPV vaccine was significantly associated with the uptake of the HPV vaccination. Therefore, awareness creation and behavior change education are mandatory to scale up the vaccination.