Background: Infection with the hepatitis B virus (HBV) is a worldwide public health issue. Egypt is classified as an area of intermediate endemicity. Hepatitis B has high materno-fetal transmission. Infants who are infected through their mothers are at a significant risk of acquiring chronic liver disease. Objective(s): To determine the prevalence and associated risk factors of HBV infection among pregnant females in Kafr El-Sheikh Governorate as well as to compare HBV serologic profile among HBV vaccinated and non-vaccinated pregnant women. Methods: A cross sectional study was performed on 456 pregnant women attending antenatal care clinics at Kafr El-Sheikh general hospitals. This study was carried out from June 2020 through February 2021. Sociodemographic data were collected through a predesigned questionnaire. Study participants were screened for hepatitis B core antibody (anti-HBc), hepatitis B surface antibody (anti-HBs) and hepatitis B surface antigen (HBsAg). Positive ones for the latter were subjected to hepatitis B e antigen (HBeAg) detection. Results: The majority of pregnant women (73%) were susceptible to HBV infection, while 2.4% had immunity following infection and 12.4% had immunity related to vaccination. HBsAg was detected in 1.8% and sole anti-HBc detected in 9.9%. There was a significant statistical association between HBsAg positivity and age, lack of vaccination, family history of HBV infection and unsafe injection. Conclusion: The prevalence of HBsAg among pregnant women in Kafr El-Sheikh Governorate was 1.8% especially in those with risk factors. Thus, it is highly recommended that the Ministry of Health and Population should implement a program to screen all pregnant women for HBV at the antenatal care units. In addition, hepatitis B vaccine proved to be an effective tool against HBV infection among studied pregnant women.