Human papillomavirus (HPV) is a small, non-enveloped, double-stranded DNA virus, belonging to the family of Papillomaviridae. It is a highly common infectious agent, which causes one of the most widespread sexually transmitted infections (STIs), involving approximately 20% of sexually active female adolescents and up to 80% of female adults aged 50years. There exist two major preventative strategies: namely, anti-HPV vaccination and cervical screening. Healthcare providers, including nurses, can play a crucial role in HPV immunization campaigns, counteracting vaccine hesitancy and doing advocacy and counseling. To explore the overall knowledge of HPV and HPV-related issues, as well as the attitudes and practices of Israeli nurses, a modified and adapted, previously validated knowledge, attitudes, and practices (KAP) questionnaire was administered to a sample of 565 participants, 481 female (85.1%) and 84 male (14.9%). Most of them were married (n = 428, 75.8%), with more than 5years of experience (n = 405, 71.7%), working in the general/internal medicine ward (n = 432, 76.5%), and Jewish (n = 352, 62.3%). Only 87 nurses (15.4%) got vaccinated against HPV. Forty-four (9.1%) of the female nurses never underwent a Pap smear. Higher percentages of right replies were reported only for the item related to available preventative strategies for cervical cancer (82.1% versus 17.9%). Higher percentages of wrong replies were reported for items related to body regions that can be infected by HPV (60.5% versus 39.5%), percentage of girls aged 15-18years being sexually active (68.0% versus 32.0%). Similar percentages of right and wrong replies were reported for items related to whom the Pap test is recommended (52.7% versus 47.3%), Israeli Ministry of Health's guidelines for anti-HPV vaccination (50.4% versus 49.6%), and percentage of boys aged 15-18years being sexually active (44.4% versus 55.6%). Concerning the reliability of the third section of the questionnaire, Cronbach's alpha was deemed to be acceptable (α = 0.64). Statistically significant determinants of reporting lower scores in HPV-related attitudes and practices were religion (OR 1.44 [95%CI 1.02-2.04]), and male offspring (OR 1.22 [1.03-1.44]). This study has important implications for policy- and decision-makers in that they should be aware of the overall poor and unsatisfactory level of HPV-related knowledge among Israeli nurses and implement multipronged HPV vaccine promotion programs, taking into account the challenges of a multicultural and diverse society like Israel.