Heterogeneity of hepatitis B virus C-gene sequences: implications for amplification and sequencing.

Abstract:

:Occasionally direct sequencing of amplified hepatitis B virus DNA leads to weak signals on autoradiograms. Using amplified C-gene sequences we investigated whether this is due to sequence heterogeneity of virus populations and use of inappropriate primers for direct sequencing. High C-gene sequence heterogeneity (point mutations, stop codons and a one codon deletion) was observed in HBV genomes from serum of a chronic carrier who underwent interferon treatment. The type of C-gene mutations detected by direct sequencing depended on the type of primers used. Cloning and sequencing of amplified C-gene sequences demonstrated that this was due to mutations in the region complementary to the sequencing primer. These data demonstrate the existence of novel HBV C-gene mutants and imply that multiple or degenerate sequencing and amplification primers are essential for accurate evaluation of the extent of HBV C-gene heterogeneity. Based on comparative sequence analysis of all available completely or incompletely sequenced C-genes, guidelines for optimal primer design are proposed for similar studies.

journal_name

J Hepatol

journal_title

Journal of hepatology

authors

Miska S,Günther S,Vassilev M,Meisel H,Pape G,Will H

doi

10.1016/s0168-8278(05)80009-1

subject

Has Abstract

pub_date

1993-04-01 00:00:00

pages

53-61

issue

1

eissn

0168-8278

issn

1600-0641

pii

S0168-8278(05)80009-1

journal_volume

18

pub_type

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