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Objective: The aim of this study was to understand the prevalence and potential risk factors of Helicobacter pylori (H. pylori) infection in Hainan Province, China. Methods: We conducted this study in 21 health service stations in 5 cities of Hainan Province from August 2022 to April 2023. We selected the various participants based on a stratified whole-group sampling method. The 14C-UBT was used to analyze H. pylori infection in 3632 participants. We also analyzed the possible relationship between variables and H. pylori infection based on chi-square test and multifactorial logistic regression. The model was evaluated by performing a Hosmer–Lemeshow goodness-of-fit test and plotting receiver operating characteristic(ROC) curves. Results: In total, the results of 3632 eligible participants (age: 14 to 93 years) were included in the analysis. The total prevalence of H. pylori infection in Hainan Province was approximately 38.7%. The prevalence of H. pylori infection was found to increase with age, stabilized in the age group of 45 to 64 years, but peaked in the age group of 65 years and older. In multifactorial analysis, the prevalence of H. pylori infection was positively associated with middle-aged adults (45–64 years), older adults (≥ 65 years), drinking, farmers, natural labor, routinely share utensils, have habit of frequent betel nut consumption, upper gastrointestinal symptoms, and family history of gastric cancer. The factors negatively associated with prevalence included family size ≤ 3, washing hands often before meals, frequent exercise, regular meals, and frequent consumption of fruits and vegetables. In addition, the Hosmer–Lemeshow test showed a good fit (χ2 = 12.983, P = 0.112) and the area under ROC was 0.631 (95%CI: 0.613 ~ 0.649). Conclusion: The prevalence of H. pylori infection in Hainan Province was observed to be moderate and closely related to age, local socioeconomic conditions, hygienic status and dietary habits. [ABSTRACT FROM AUTHOR]