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Background: Hepatitis E virus (HEV) is an emerging pathogen associated with endemic and acute viral hepatitis. In this study, we investigate the HEV seroprevalence and putative risk factors by a nationwide cross-sectional study in the Republic of Korea. Methods: The prevalence of anti-HEV antibody was investigated in 2,450 serum samples collected in fourth Korea National Health and Nutrition Examination Survey. In addition, epidemiological information on possible risk factors including gender, age, education, occupation, and residence location for exposure to HEV was obtained. Results: The frequency of anti-EIA reactive sample was 5.9% (144/2450). The individuals in groups with male, older age, low education level and living in rural or coastal regions had high seroprevalence estimates (P ≤ 0.001). In addition, seroprevalence was significantly higher among individuals with self-identified skilled agricultural, forestry, and fishery workers (31.3%, P < 0.001). Conclusions: This study provides valuable data that could be used to investigate associations of HEV seroprevalence and putative risk factors by a nationwide cross-sectional study. The high HEV seroprevalence of skilled agricultural, forestry, and fishery workers and individuals lived in coastal and rural area indicated that zoonotic transmission is an important risk factor for HEV infection in the republic of Korea. Further studies that include detailed and continuous nationwide surveys are required to identify unrecognized risk factors and to monitor the HEV infection prevalence. [ABSTRACT FROM AUTHOR]