Analysis of hepatitis B virus drug-resistant mutant haplotypes by ultra-deep pyrosequencing

Sun-Young Ko, H. B. Oh, C. W. Park, H. C. Lee, J. E. Lee

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Direct sequencing and reverse hybridization are currently the main methods for detecting drug-resistance mutations of hepatitis B virus (HBV). However, these methods do not enable haplotype analysis so they cannot be used to determine whether the mutations are co-located on the same viral genome. This limits the accurate identification of viral mutants that are resistant to drugs with a high genetic barrier. In our current study, ultra-deep pyrosequencing (UDPS) was used to detect HBV drug-resistance mutations in 25 entecavir-treated and five treatment-naive patients. Of the 25 entecavir-treated patients, 18 had experienced virological breakthrough and two exhibited reduced susceptibility to entecavir. The results obtained by UDPS were compared with those of direct sequencing, and the haplotypes of the drug-resistant HBV mutants were analysed. The average number of reads per patient covering the region in which drug-resistance mutations are located was 1735 (range 451-4526). UDPS detected additional drug-resistance mutations not detected by direct sequencing in 19 patients (mutation frequency range 1.1-23.8%). Entecavir-resistance mutations were found to be co-located on the same viral genome in all 20 patients displaying virological breakthrough or reduced susceptibility to entecavir. In conclusion, UDPS was not only sensitive and accurate in identifying drug-resistance mutations of HBV but also enabled haplotype analysis of the mutants. This method may offer significant advantages in explaining and predicting the responses of patients with HBV to antiviral therapy.

Original languageEnglish
JournalClinical Microbiology and Infection
Volume18
Issue number10
DOIs
Publication statusPublished - 2012 Oct 1
Externally publishedYes

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Hepatitis B virus
Haplotypes
Drug Resistance
Mutation
Pharmaceutical Preparations
Viral Genome
Mutation Rate
Antiviral Agents
entecavir
Therapeutics

Keywords

  • Drug-resistance mutation
  • Haplotype analysis
  • Hepatitis B virus
  • Ultra-deep pyrosequencing

ASJC Scopus subject areas

  • Microbiology (medical)
  • Infectious Diseases

Cite this

Analysis of hepatitis B virus drug-resistant mutant haplotypes by ultra-deep pyrosequencing. / Ko, Sun-Young; Oh, H. B.; Park, C. W.; Lee, H. C.; Lee, J. E.

In: Clinical Microbiology and Infection, Vol. 18, No. 10, 01.10.2012.

Research output: Contribution to journalArticle

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