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Background: This study assessed awareness, attitudes, and uptake of human papillomavirus (HPV) vaccination and cervical cancer screening in detained women. Methods: The cross-sectional study was conducted from April to June 2022 in four women prisons in Italy. Results: 41.1% of participants recognized HPV infection as an sexually transmitted diseases (STD), 36.4% identified cervical, and 16.8% oral cancer as an HPV-associated disease. Overall, 70% had never heard of HPV vaccination, and 45.8% believed it is effective to prevent cervical cancer. Among the age-eligible women for HPV vaccination, none reported to have undergone it, nor had talked about it with a physician in the previous year. Only 13.5% declared to have ever undergone cervical cancer screening, and adherence was significantly higher in those who were involved in a working activity in prison, who were aware that HPV infection is an STD and that can cause cervical and oral cancer, and who were older at their first sexual intercourse. Conclusion: These findings documented an extremely low awareness of HPV infection and an unsatisfactory adherence to prevention through HPV vaccination and cervical cancer screening. There is a need for evidence-based interventions for incarcerated women to promote participation in HPV vaccination and cervical cancer screening programs as routine activities.