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Reportedly transmitted through unprotected sexual intercourse with infected person(s), experts have estimated new cases of hepatitis B virus (HBV) infections to be over 70,000 per year in the United States. With little or no records of such in Nigeria, this study investigated the prevalence of hepatitis B surface antigen at different trimesters of pregnancy in women who visit general hospital Agbor, Delta state, Nigeria; for antenatal care. A total of one hundred (100) pregnant and fifty (50) non-pregnant (control) women were ethically recruited for the exercise. They were then sub-grouped by age and duration of pregnancy (trimester); and an Acon serological strip was used to obtain blood samples from each subject. Obtained blood was then assayed for the presence of hepatitis B virus (in serum) and compared with those of control (non-pregnant) group. Following careful comparison of differences in mean (using the Analysis of variance), study found a 5% prevalence rate (of hepatitis B) in pregnant than non-pregnant (2% prevalence) women. Study also observed a statistically significant increase in hepatitis B surface antigen for non-pregnant women of age bracket 20-24 years (2.6% prevalence) to pregnant women of between 20-24 years. HBV infection therefore has high prevalence rate in pregnant than non-pregnant women as they are often more exposed to unprotected sexual intercourse. We recommend regular and continuous HBV screening in pregnancy to help circumvent HBV infection related ailments and complications. The same is also suggested for non-pregnant women for purpose of proper vaccination.