Direct detection of HBV preC mutants in heterogeneous viral populations by a modified DNA sequencing method.
A modified method for the direct sequencing of double-stranded DNA products of PCR amplification is described and has been applied to the analysis of hepatitis B virus (HBV) preC/C region in samples from persistently infected patients with chronic hepatitis. Data was obtained from both hepatitis B e antigen (HBeAg)-positive and -negative chronic carriers. A high prevalence of mixed viral populations (wild-type genomes and mutated sequences with a TAG stop codon in the distal preC region at position 1895-1897) was shown in the HBeAg-positive group; a homogeneous (either mutated or wild-type) viral population was detected in all but one of the long-term HBeAg-negative, untreated chronic carriers, thus suggesting that pre-core mutants can be rapidly generated and selected during the natural course of HBV infection.
Manzin
A
A
Institute of Microbiology, University of Ancona, Italy.
Paolucci
S
S
Lampertico
P
P
Menzo
S
S
Rumi
M G
MG
Colombo
M
M
Clementi
M
M
D001483:Base Sequence; D002908:Chronic Disease; D014644:Genetic Variation; D006509:Hepatitis B / Q000235:genetics*; D006512:Hepatitis B Core Antigens / Q000235:genetics*; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods
D006512:Hepatitis B Core Antigens; D011498:Protein Precursors