Objectives The aims of this study were to evaluate the long-term seroprotection after hepatitis B virus (HBV) vaccination and to assess the need for a booster dose among children vaccinated during infancy.Patients and methods A cross-sectional study was carried out in two areas in the Beni-Suef governorate (El-Wasta city and Blevia village) using a multistage random sampling. Face-to-face interviews of 362 children and their parents were carried out. Blood samples were collected to assess hepatitis B surface antigen (HBsAg), total hepatitis B core antibody (anti-HBc), and antibodies to hepatitis B surface antigen (anti-HBs) using ELISA. Positive samples for HBsAg/anti-HBc were subjected to quantitative HBV-DNA detection by a real-time PCR. Children with HBV non-sero-protection received a booster dose of the HBV vaccine to assess their anamnestic response 4 weeks later.Results Four (1.1%) of the participants had evidence of breakthrough HBV infection and the seroprotection rate was 57%. On multivariate analysis, the risk of non-seroprotection was significantly higher among children aged 5to less than 10 years, 10 to less than 15, and at least 15 years compared with younger children [adjusted odds ratio (AOR) = 5.2, 18.6, and 19.7, respectively] and those living in rural compared with urban areas (AOR = 2.1). Out of 117 children with non-sero-protective levels, 86.3% developed an anamnestic response after a booster dose.Conclusion Long-term HBV immunity exists among children vaccinated during infancy even in case of reduced or absent anti-HBs over time. The high anamnestic response rate developed signifies the presence of immune memory and indicates that there is no need for a booster dose.