Natural populations of woodchuck hepatitis virus contain variant precore and core sequences including a premature stop codon in the epsilon motif.

We have determined a consensus sequence and the type and the frequency of spontaneous sequence variations in the woodchuck hepatitis virus (WHV) precore gene and the 5' region of the core gene in 101 serum samples from 53 naturally WHV-infected woodchucks by polymerase chain reaction sequencing. Twenty of the 53 woodchucks were found to have variant sequences. Ten patterns of variant sequences were identified in these 20 animals. WHV sequences from 4 woodchucks had 1 nucleotide change, 3 had 2 nucleotide changes and 3 had 3 nucleotide changes. The nucleotide changes were not randomly distributed, but were limited to only 8 sites. Four sites were in the epsilon motif of the precore gene and four were in the 5' region of the core gene. Sixteen of the 53 (30%) woodchucks had precore sequence variants. All altered sites were analogous to previously described mutations in hepatitis B virus. There was a nucleotide change at nucleotide 2016 in codon 29 of the precore region that produced a stop codon in 4 animals. This site is analogous to a common hepatitis B virus e antigen mutation. The sequence from the initial blood samples from 3 of 4 animals with this stop codon producing variant appeared to be the consensus sequence; however, in later samples the variant occurred as a mixed infection with the consensus sequence. The mixed infections were chronic and the proportion of the variant sequence was maintained or increased in the course of infection. In the fourth animal only the variant was found and it persisted for over 14 months of infection. WHV appears to be a valuable model for the study of the structure and function of the hepadnavirus precore region.