A NOVEL METHOD FOR EFFICIENT AMPLIFICATION OF WHOLE HEPATITIS-B VIRUSGENOMES PERMITS RAPID FUNCTIONAL-ANALYSIS AND REVEALS DELETION MUTANTS IN IMMUNOSUPPRESSED PATIENTS
S. Gunther et al., A NOVEL METHOD FOR EFFICIENT AMPLIFICATION OF WHOLE HEPATITIS-B VIRUSGENOMES PERMITS RAPID FUNCTIONAL-ANALYSIS AND REVEALS DELETION MUTANTS IN IMMUNOSUPPRESSED PATIENTS, Journal of virology, 69(9), 1995, pp. 5437-5444
Current knowledge of hepatitis B virus (HBV) sequence heterogeneity is
based mainly on sequencing of amplified subgenomic HBV fragments, Her
e, we describe a method which allows sensitive amplification and simpl
ified functional analysis of full-length HBV genomes with or without p
rior cloning. By this method, a large number of HBV genomes were clone
d from sera of six immunosuppressed kidney transplant patients, Two si
ze classes of HBV genomes, one 3.2 kb and another about 2.0 kb id size
, were found in all patients. The genome population from one serum sam
ple was studied in detail by size analysis of subgenomic PCR fragments
and sequencing. Regions with deletions and insertions were mapped in
the C gene and pre-S region, UP to 100% of HBV genomes in all other im
munosuppressed patients also had deletions in the C gene, Our results
demonstrate the potential Of the established method for the structural
and functional characterization of heterogeneous populations of compl
ete virion-encapsidated HBV DNAs and suggest that HBV genomes with C g
ene deletions can have a selective advantage in immunosuppressed patie
nts.