BACKGROUND: Human Papillomavirus (HPV) infection has garnered significant attention due to its high prevalence and association with various cancers and other health conditions. Composite lifestyle factors may influence the risk of HPV infection, yet their cumulative impact remains insufficiently explored. This study aims to explore the association between the Life's Essential 8 (LE8) Score and HPV infection status, highlighting the potential role of lifestyle and health behaviors in HPV infection prevention. METHODS: Utilizing data from the National Health and Nutrition Examination Survey (NHANES) spanning 2005-2016, we analyzed the health and nutritional statuses of 6,773 participants after excluding those with missing HPV infection status, inability to calculate the LE8 Score, and missing covariate data. The LE8 Score was computed based on eight cardiovascular health metrics, encompassing both health factors (BMI, non-HDL cholesterol, blood pressure, and blood glucose) and health behaviors (physical activity, diet, sleep duration, and nicotine exposure). HPV infection status was determined through vaginal swab specimens analyzed using various Roche assays. Multivariate logistic regression, the restricted cubic splines (RCS) analysis and weighted quantile sum (WQS) regression were employed to assess the association between LE8 Score and HPV infection risk. RESULTS: Our findings indicate a significant inverse association between the LE8 Score and HPV infection risk. Participants with medium and high LE8 Scores exhibited a 21% and 31% lower risk of HPV infection, respectively, compared to those with low LE8 Scores in multivariate logistic regression models. The analysis also revealed that lifestyle factors, particularly nicotine exposure and blood pressure, significantly contributed to the observed association. CONCLUSION: The study underscores the importance of healthy lifestyle behaviors in reducing the risk of HPV infection. Public health strategies promoting such behaviors could complement existing HPV prevention measures, potentially lowering the burden of HPV-related diseases. Future research should further investigate the mechanisms underlying this association and the effectiveness of lifestyle interventions in diverse populations.