Phylogenetic Analysis of Hepatitis B Virus among Household Members with HBV Chronic Infection

Intrafamilial spread of Hepatitis B virus (HBV) infection in Iran has only been investigated with serological testing without using molecular studies as the most informative and definitive type of analysis. In the present study, intrafamilial transmission of HBV among family members of Iranian index HBsAg carriers was investigated using phylogenetic analysis of the S region of the viral genome. Nested polymerase chain reaction was used for detection of HBV DNA in serum samples from 22 index and 43 contact patients with chronic HBV infection. HBV DNA was detected in 37 samples (14 indexes, 23 contacts). The S gene region of the DNA isolates was subjected to direct sequencing and phylogenetic analysis by Bioedit, Mega and Phylip programs. All isolates (from 26 patients) were clustered with genotype D, of which 24 strains were of subgenotype D1, subtype ayw2, while 2 additional strains were of subgenotype D2, subtype ayw3. Evidence of intrafamilial transmission of the virus was found in 8 families studied phylogenetically. Overall, 60 changes were detected in the amino acid sequences of the surface antigen protein in 23 patients. Four premature stop codons occurred in 3 isolates at residues 69 and 182. Seven out of 8 families displayed 25−100% common amino acid substitutions among their members. Our data corroborated intrafamilial transmission of HBV, as evidenced by concordant HBV genotype among household members, viral sequence homology and close genetic relatedness of the strains on the phylogenetic tree, and horizontal transmission of S gene mutations among family members.