Background: Hepatitis B Virus infection is one of the world's most common viral hepatitis and a significant global public health issue with varied prevalence across the world. Aim of the research was to investigate the effectiveness of a structured teaching program on pregnant women' knowledge and attitude regarding hepatitis B virus infection with pregnancy. Design: A quasi-experimental research design was adopted. Setting: the research was performed at an outpatient clinic for obstetrics and gynaecology affiliated to the Benha University Hospital. Sample: A purposive sample of 220 pregnant women were recruited. Tools: three tools were used for data collection; a structured self-administered questionnaire, knowledge assessment sheet and modified likert scale for maternal attitude. Results: illustrated that there was a highly statistically significant improvement in relation to pregnant women’ knowledge and attitude regarding hepatitis B virus infection at post-intervention phase compared to pre-intervention phase (P ≤ 0.001). As well, there were a positive highly statistically significant correlation between total knowledge and total attitude scores at preintervention and post-intervention phases (P ≤ 0.001). Conclusion: the structured teaching programme had a positive effect on the improvement of pregnant women' knowledge and attitude regarding hepatitis B virus infection with pregnancy. Recommendations: Educational program about hepatitis B virus infection should be a part of antenatal care for every pregnant woman.