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Zufan Asaye,1 Temesgen Aferu,2 Adane Asefa,3 Desalegn Feyissa,2 Tolcha Regasa,2 Oliyad Kebede,2 Diriba Feyisa,2 Mulugeta Feyisa4 1Department of Statistics, College of Natural Sciences, Mizan-Tepi University, Mizan-Aman, Ethiopia; 2School of Pharmacy, College of Health Sciences, Mizan-Tepi University, Mizan-Aman, Ethiopia; 3School of Public Health, College of Health Sciences, Mizan-Tepi University, Mizan-Aman, Ethiopia; 4Department of Midwifery, College of Health Sciences, Selale University, Fiche, EthiopiaCorrespondence: Temesgen AferuSchool of Pharmacy, College of Health Sciences, Mizan-Tepi University, Mizan-Aman, EthiopiaEmail temesgenaferu@gmail.comBackground: Hepatitis B virus(HBV) infection is a global public health problem, even though its prevalence is disproportionately high in low- and middle-income countries. Mother-to-child transmission is a major route of HBV transmission in endemic areas. This study aimed to assess the prevalence of HBV and its determinants among pregnant women attending antenatal care at Mizan-Tepi University Teaching Hospital and Mizan Health Center, Southwest Ethiopia.Methods: A cross-sectional study was conducted between January 13th 2020 and February 5th 2020 among 370 pregnant women. The sample size was proportionally allocated to each health institution according to the total pregnant women on antenatal care at the respective health institution and a consecutive sampling technique was used to select study participants. Serum hepatitis B surface antigen (HBsAg) was tested using a rapid diagnostic test. Data were analyzed using Statistical Package for Social Sciences (SPSS) software version 22. Multiple logistic regression analysis was done to identify the independent predictors of HBsAg serostatus at p-value < 0.05.Results: Three hundred seventy of the total 375 pregnant women participated in the study resulting in a response rate of 98.7%. Twenty-two (5.9%) of the pregnant women screened were found positive for HBsAg (prevalence=5.9%; 95% CI: 3.9– 8.80%). History of contact with jaundice patients (AOR=9.87; 95% CI: 2.98– 32.65), sharing sharp materials (AOR=3.96; 95% CI: 1.23– 11.08) and history of multiple sexual partners (AOR=6.77; 95% CI: 2.44– 18.78) were significantly associated with Hepatitis B Virus infection.Conclusion: The endemicity of hepatitis B virus seroprevalence is intermediate in the study settings. Factors associated with hepatitis B virus serostatus were behavioral; hence, modification of these factors may help to prevent the infection.Keywords: hepatitis, hepatitis B, hepatitis B surface antigen, pregnant women