Hepatitis B virus infection is endemic in sub-Saharan Africa, and accounts for a significant proportion of morbidities and mortalities in Ghana. Infection with HBV during pregnancy can result in life-threatening complications to both mother and child. To improve their quality of life, the free maternal care was introduced to grant pregnant women cost-free access to antenatal and postnatal services. The study analysed the prevalence of HBV infection among pregnant women receiving free antenatal care in a tertiary hospital in Ghana. This was a retrospective cross-sectional study, where secondary data of pregnant women who accessed free antenatal services at the Trafalga hospital, Ho, from 2016 to 2017 were retrieved from the hospital’s database. Data on hepatitis B surface antigen reactivity test, age and period of turnout were analysed with Microsoft Excel and Graph pad prism version 6. A total of 2,634 pregnant women assessed free antenatal care from January 2016 –December 2017, with 10% rise in turnout in 2017. The age of the study population was fairly young, ranging from 13–52 years and mean of 29.87±5.83. The prevalence of HBV infection among pregnant women in the entire study was estimated to be 6.0%, while that of 2016 and 2017 were 5.3% and 6.7% respectively. Turnout for antenatal services peaked in July, which also recorded the highest prevalence of HBV infection among the pregnant women. Our study, first of its kind show an HBV prevalence of 6.0% among a large population of pregnant women who accessed free antenatal services at a tertiary hospital in Ghana. The study evaluates the influence of the free maternal care policy on antenatal attendance and HBV infection rates among pregnant women.