On the basis of a seroepidemiological survey of hepatitis B virus (HBV) infection conducted on 6208 random serum samples from four provinces of Thailand, we found 19 of 246 (7.7%) hepatitis B surface antigen (HBsAg)-positive samples with unusual serological constellations of HBV infection. Ten samples tested positive for HBsAg, anti-HBc (anti-hepatitis B core antibody), and anti-HBs (anti-hepatitis B surface antibody) markers (group I), 3 specimens were HBsAg and anti-HBs positive without detectable anti-HBc (group II), and the remaining 6 specimens showed only HBsAg (group III). In group I, 7 of 10 HBsAg-positive sera could be confirmed by HBsAg neutralization, yielding positive results for all samples. None of the group II sera were available in sufficient amounts for confirmation. In group III, five of six sera were confirmed by HBsAg neutralization, with four showing a positive reaction. HBV DNA was detected in 7 of 10 (70%) specimens in group I, in 1 of 3 (33.3%) specimens in group II, and in 3 of 6 (50%) specimens in group III. On the basis of HBsAg neutralization, HBV DNA was found in five of seven (71.4%) HBsAg-positive samples in group I and in three of four (75%) HBsAg-positive samples in group III, whereas the one confirmed HBsAg-negative sample in group III also remained negative for HBV DNA. Amino acid sequences were compared with those specifying the "a" determinant of the wild-type virus, particularly focusing on HBV-S protein variations between positions 110 and 160. Among 11 HBV DNA-positive sera, G145A was detected in 2 samples in group I, with the remaining samples identical to the wild-type virus. These unusual serological profiles may be due to the altered immune response of the host or to HBV variants.