The risk of vertical transmission of hepatitis B virus (HBV) varies with type of viral endemicity, degree of maternal infection and genomic characteristics of the virus. The aim of this study is to estimate this risk in Tunisia using serological and molecular methods to evaluate HBV replication, to determine viral genotypes and to detect presence of occult hepatitis in 2709 pregnant women. Serological markers were detected by ELISA methods, Genotype was determined by PCR-RFLP and occult hepatitis by nested-PCR. Four percent of women were positive for HBsAg; only 3% of them were also positive for HBeAg. Viral replication, over than 10(3) copies/ml, was detected in 61% of positive HBsAg patients. Three viral genotypes were detected: D (95%), B (3%) and A (3%). Occult hepatitis was detected in 4% of sera with "anti-HBc isolated" profile. In conclusion, the risk of vertical transmission of HBV exists in Tunisia. It increases by frequency of precore mutants, predominance of the genotype previously associated with high levels of replication and possibility of occult hepatitis B. These results show the importance of screening by serological HBV markers systematically during pregnancy with evaluation of viral replication in order to prevent vertical risk by efficient tools.