Receiving the HPV vaccine at a young age before being exposed to the virus is essential to improve HPV-related cancer prevention. In many countries in the world, like Italy, the HPV vaccine is offered free of charges to boys and girls aged 12-14. Despite this incentive, the HPV vaccine uptake in Italy remains low. Several interventions and campaigns have been implemented to educate parents and providers. However, children of 12-14 years old have been left out by prevention and education efforts. This study explored middle school children’s knowledge and perceptions of the HPV virus and vaccine with the purpose to identify gaps and misperceptions and provide recommendations to develop educational interventions. A total of nine focus groups were conducted, and discussions were analyzed along with 171 anonymous questions collected from the children. Findings included a lack of knowledge of HPV prevention guidelines, negative emotions associated with both the HPV vaccine and vaccines in general, misperceptions between prevention and care, and the desire to receive specific information.