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Background: Hepatitis B infection is a universal concern. This infection can lead to chronic liver disease and hepatocellular carcinoma. Neonates born to HBsAg-positive mothers are at high risk of chronic hepatitis B virus (HBV) infection, especially for HBeAgpositive mothers or neonates who have not received hepatitis B immunoglobulin (HBIg) and HBV vaccines. Objectives: The aim of this study was to evaluate the efficacy of post-exposure prophylaxis in these infants to prevent infection. Patients and Methods: Thirty-eight infants born to HBsAg-positive mothers between September 2006 and September 2013 were followed. The investigation evaluated whether the standard prevention protocol of neonatal HBV transmission including HBIg at birth and receiving three doses of vaccine at birth and 2 and 6 months of age was performed, followed by post-vaccination tests (evaluation of HBsAg and HBsAb titer at 9 to 18 months of age) to determine subsequent infection. HBsAb titer ≥ 10 was considered as criterion for effectiveness of the prophylaxis procedure. The acquired data were analyzed using SPSS software (Version 18). The results are reported in descriptive tabulations. Results: Ninety seven percent (97%) of infants received HBIg at birth in the hospital. Generally, all of them received the first, second and third doses of vaccine at birth, 2 months, and 6 months after birth, respectively. Information for 35 mothers infected with HBV and 38 infants was available. The mean age of the mothers was 30.3 years. The results indicated that 20% of mothers were HBeAg positive. HBsAg was positive in one (2.6%) infant born to an HBeAg-positive mother. Around 94% of infants’ HBsAb titers were ≥ 10, and 5.8% were reported as non-responders. Conclusions: The vertical transmission prevention program used in the study population in Tehran, which had an appropriate sample size, is effective. Additional doses of the vaccine can be useful in raising the effectiveness of immunoprophylaxis for infants at high risk of HBV infection. Also, emphasis must be set on post-vaccination testing. [ABSTRACT FROM AUTHOR]