Implementing a HPV vaccination program is currently under evaluation by the Turkish health ministry. For screening and vaccination programs to be successful, the cooperation of nurses is essential. We aimed to evaluate (1) basic knowledge of nurses and nursing interns regarding HPV infection and cervical cancer, (2) their attitudes towards smear testing and HPV vaccination, and (3) their viewpoint on vaccination of school age children. This cross-sectional study was undertaken at Bezmialem Vakif University. The survey was designed to assess knowledge about HPV infection, Pap smear testing, cervical cancer, HPV vaccine, attitudes towards HPV vaccination, and school-based vaccination programs. Validity content was determined by expert gynecologists, and a pilot study was performed on 10 nurses. A total of 550 questionnaires were handed out; 499 were completed. Our response rate was 90.7%. Fifty-nine participants answered all the knowledge questions correctly. The calculated knowledge score of the female participants was 6.99 ± 2.22, the male participants was 5.89 ± 2.92. Female participants were more knowledgeable (p < 0.0001). Out of the 353 female participants, 18.6% (n = 66) had undergone smear testing. There were 20 (5.6%) female and 6 (4%) male participants who were vaccinated against HPV. The leading answer for not having a HPV vaccine was "I'm not at risk for a HPV infection" (n = 106, 34.9%). There was a statistical relationship between "HPV knowledge score" and answering "Yes" to "Do you want your children/future children to be vaccinated? (p = 0.001) and "Do you think including the vaccine in the Turkish immunization program is necessary?" (p = 0.001). Nurses in our cohort seem to have satisfactory basic knowledge regarding HPV infection; however, their viewpoints on vaccination were not favorable. Strategies and intervention materials for HPV vaccination will be necessary if a national immunization program will be initiated.