A mass hepatitis B vaccination program began in Taiwan in 1984. In order to determine the immune status of hepatitis B virus (HBV) infection among preschool children, a total of 25 kindergartens in 20 townships and metropolitan precincts in central Taiwan were randomly selected through stratified sampling. Serum specimens of 2130 healthy preschool children aged 2-6 years old were screened for the HBV markers and liver function in 1996. HBV surface antigen (HBsAg), antibody against HBsAg (anti-HBs) and antibody against HBV core antigen (anti-HBc) were tested by reverse passive hemagglutination (RPHA), enzyme immunoassay (EIA) and radioimmunoassay (RIA) using commercial kits. HBV vaccination rate of the preschool children was 98%, and complete vaccination rate (three or four doses of HBV vaccine) was 94%. The HBsAg seropositive rate was 4.5% among incomplete vaccinees and 1.3% among complete vaccinees. The anti-HBs was detectable in 1637 of 2000 complete vaccinees (81.9%) and in 53 of 88 incomplete vaccinees (60.2%). The overall prevalence rate of anti-HBc was 2.4% (52 of 2130). The older the age, the lower the anti-HBs seropositive rate. The anti-HBs seropositive rats for complete vaccinees were 100% at 2 years old and 75% at 6 years old. There were no significant differences in HBsAg-seropositive rates and anti-HBs-seropositive rates among different residential areas or ethnic groups. There were three children who were seropositive on HBsAg, anti-HBs and anti-HBc, whether they were infected by the vaccine-induced escape mutant of HBV deserves scrutiny.