ObjectiveThis study assesses the knowledge, practices, educational needs and hospital engagement in IPC among healthcare workers (HCWs) and students in Italy.MethodsA national cross-sectional observational study was conducted from April to July 2024. Participants completed a validated questionnaire based on WHO and CDC guidelines. Four outcomes were evaluated: (i) knowledge of IPC, (ii) self-reported compliance, (iii) satisfaction with education and (iv) hospital engagement in IPC.ResultsAmong respondents (n = 1063, 67.75% female) significant gaps in IPC knowledge were noted, with only 66.1% correctly identifying key hand hygiene moments. Compliance was low, with only 43.3% always adhering to hand hygiene rules. Satisfaction with IPC education was also low (28%). While alcohol-based hand gel was widely available (84.2%), only 36.6% received formal hand hygiene training before onboarding.Multivariate analysis revealed that female HCWs had higher IPC knowledge [adjusted odds ratio (aOR) 1.55, 95% CI: 1.04-2.31] and better compliance (aOR 1.72, 95% CI: 1.01-2.92). Younger HCWs demonstrated greater knowledge and satisfaction. Surgical ward staff, despite greater education satisfaction, exhibited lower IPC knowledge (aOR 0.53, 95% CI: 0.32-0.87). Nurses and health assistants received more hospital IPC training (aOR 1.59, 95% CI: 1.04-2.46; aOR 5.83, 95% CI: 1.94-18.89, respectively). Finally, the presence of a hospital IPC team positively correlated with all outcomes, including higher IPC knowledge (aOR 2.68, 95% CI: 1.79-4.05) and adherence (aOR 3.4, 95% CI: 1.92-6.3).ConclusionThe study reveals substantial gaps in IPC knowledge, practices and education among Italian HCWs, emphasizing the urgent need for effective targeted interventions that require robust institutional support.