Background: The human papillomavirus (HPV) is one viral infection of the reproductive system that causes cervical cancer in women, however; less emphasis is placed on the level of uptake/practice of Human Papillomavirus vaccination and associated factors; appropriate information is required for an efficient immunization program. Objectives: This study aimed to assess the uptake of Human Papillomavirus vaccination and associated factors among female primary school students in Hawassa City, Sidama Region, Ethiopia, 2022. Methods: A school-based cross-sectional study was conducted, across 660 respondents selected by two-stage sampling techniques from June 16/2022 to July 16/2022. The data was collected by using face-to-face interviews and entered into Epi data version 4.6 then exported to SPSS version 26 for analysis. Descriptive variables were computed using a measure of central tendencies & reported by the frequency with percentage. The factors influencing students' uptake of the HPV vaccine were assessed using logistic regression analysis. Variables with a P-value of <0.25 in bivariable logistic regression were candidates for multivariable and those with a P-value of <0.05 in multivariable logistic regression were considered a statistically significant factor for uptake of the Human Papillomavirus vaccine. Result: A total of 660 study participants with 100% response rate were included in the study. Overall, 59.4% (95%, CI: 55.6%-62.9%) were Uptake of the HPV vaccine among female primary school students. The Uptake of HPV vaccine were significantly associated with students age (AOR=1.475; 95% CI=1.036-2.098; P=0.03), also being students family monthly income >4000ETB (AOR=2.397; 95% CI=1.316-4.368; P=0.04), Good knowledge towards HPV vaccine (AOR=1.623; 95% CI=1.078-2.482; P=0.020), Positive Attitude towards HPV vaccine (AOR=1.739; 95%,CI=1.197-2.526), and Family/guardian support to get HPV vaccination (AOR=4.306, 95% CI=2.986-6.209; P=0.000) . Conclusion and Recommendations: Students' uptake of the HPV vaccine was encouraging (59.4%), however, to increase student Human Papillomavirus vaccine uptake; Families, health care workers, and schools should work together on the knowledge and attitude of the community, that is effective public health strategies to improve the uptake of HPV vaccination.