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INTRODUCTION: Perinatal transmission is the dominant mode of transmission of the hepatitis B virus in sub-Saharan Africa. The purpose of the present study was to determine the prevalence of HBsAg positivity in pregnant women attending prenatal clinics, and to identify the associated factors. METHODS: It was a descriptive and analytical cross-sectional study, with data collection over a period of one month at the Obstetrics and Gynecology Department (CUGO) at Hubert Koutoukou Maga National Hospital Center (CNHU-HKM) affiliated with the Medical School in Cotonou, Benin. The study population is all pregnant women of all gestational age who received prenatal care at the CUGO during the study period. Pregnant women who signed a consent form were screened using the Determine® Ag HBs rapid diagnostic test. The Pearson Chi-square statistical test was used for comparisons (p<0.05). RESULTS: Of the 250 pregnant women cared for during the study period, 166 consented to the screening. The prevalence of carriage of HBsAg was 14%. Anti-HBc and HBeAg were negative in all cases, indicating chronic infection. A history of hepatic pathology (p=0.01) and the presence of HIV infection (p=0.01) were significantly associated with the carriage of HBs Ag. CONCLUSION: The frequency of HBsAg carriage was high in pregnant women. It is essential to set up a systematic vaccination program, in the first hours of life, for all newborns in our country.