To study sexual transmission of hepatitis D virus (HDV), 52 spouses of 56 index patients were observed and HDV genomes from antibody to HDV (anti-HDV)-positive couples were sequenced. Of the spouses, 11 (21%) were serum HBsAg positive, 3 (27%) of whom were also anti-HDV positive. The HDV sequences between spouses were found to be nearly identical (98% to 98.8%) in the region analyzed (nt 911 to nt 1260). Only one couple showed an identity > 90% with the genotype I HDV strains. The HDV sequences of the remaining two couples showed > 95% identity with each other and > 91% homology with genotype II, but they shared only a 73.1% to 73.7% homology with those of the first couple. The regions corresponding to the autocatalytic cleavage sites, the junction between the middle and the carboxyl terminal one-third domains, and the middle domain of the open reading frame for delta antigen on the antigenomic HDV RNA were more conserved with < 19% divergence among the three couples. Interestingly, there was a 56% divergence in the region corresponding to the carboxyl end of the open reading frame for the large delta antigen on the antigenomic HDV RNA. In summary, this study provides a direct nucleotide evidence of a common source of HDV infection in each couple. Despite divergence in the viral nucleotide sequence, both genotypes I and II were found in Taiwan and were transmitted from patients with a history of prostitute contact to spouses through sexual contact.