title: Deep sequencing analysis of quasispecies in the HBV pre-S region and its association with hepatocellular carcinoma.
abstract: The association between the evolution of hepatitis B virus (HBV) quasispecies and the development of hepatocellular carcinoma (HCC) is unknown.
  
  
  
  We used deep sequencing to examine the dynamics of HBV quasispecies and their relationship to HCC development. Thirty-two chronic hepatitis B (CHB) patients with HCC (HCC group) and 32 matched CHB patients without HCC (controls) were recruited. Fourteen patients from each group had serial sera available up to 9 years before the time of the present study. Deep sequencing of the HBV pre-S regions was performed. HBV quasispecies complexity, diversity, and intrapatient prevalence of pre-S deletions/mutations were analyzed.
  
  
  
  Compared with control patients, HCC patients had a significant greater quasispecies complexity (p = 0.04 at the nucleotide level), greater diversity (p = 0.004 and 0.009 at the nucleotide level and the amino acid level respectively), and a trend of greater complexity at the amino acid level (p = 0.065). HCC patients had a higher intrapatient prevalence of pre-S deletions and point mutations (at codons 4, 27, and 167) compared with the control patients (all p < 0.05). Longitudinal observation in the sera of 14 HCC patients showed that quasispecies complexity (p = 0.027 and 0.024 at the nucleotide level and the amino acid level respectively) and diversity (p = 0.035 and 0.031 at the nucleotide level and the amino acid level respectively) increased as the disease progressed to HCC.
  
  
  
  Increased HBV quasispecies complexity and diversity in the pre-S region, probably reflecting enhanced virus-host interplay, was associated with disease progression from CHB to HCC.
authors.affiliation: Department of Medicine, University of Hong Kong, Hong Kong Special Administrative Region, China.|Department of Medicine, Fourth People's Hospital of Shenzhen, Shenzhen, China.
authors.forename: An-Ye
authors.initials: AY
authors.lastname: Zhang
authors.affiliation: Department of Medicine, University of Hong Kong, Hong Kong Special Administrative Region, China.|State Key Laboratory for Liver Research, University of Hong Kong, Hong Kong Special Administrative Region, China.
authors.forename: Ching-Lung
authors.initials: CL
authors.lastname: Lai
authors.affiliation: Department of Medicine, University of Hong Kong, Hong Kong Special Administrative Region, China.
authors.forename: Fung-Yu
authors.initials: FY
authors.lastname: Huang
authors.affiliation: Department of Medicine, University of Hong Kong, Hong Kong Special Administrative Region, China.|State Key Laboratory for Liver Research, University of Hong Kong, Hong Kong Special Administrative Region, China.
authors.forename: Wai-Kay
authors.initials: WK
authors.lastname: Seto
authors.affiliation: Department of Medicine, University of Hong Kong, Hong Kong Special Administrative Region, China.|State Key Laboratory for Liver Research, University of Hong Kong, Hong Kong Special Administrative Region, China.
authors.forename: James
authors.initials: J
authors.lastname: Fung
authors.affiliation: Department of Medicine, University of Hong Kong, Hong Kong Special Administrative Region, China. danywong@hku.hk.|State Key Laboratory for Liver Research, University of Hong Kong, Hong Kong Special Administrative Region, China. danywong@hku.hk.
authors.forename: Danny Ka-Ho
authors.initials: DK
authors.lastname: Wong
authors.affiliation: Department of Medicine, University of Hong Kong, Hong Kong Special Administrative Region, China. mfyuen@hkucc.hku.hk.|State Key Laboratory for Liver Research, University of Hong Kong, Hong Kong Special Administrative Region, China. mfyuen@hkucc.hku.hk.
authors.forename: Man-Fung
authors.initials: MF
authors.lastname: Yuen
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000453:epidemiology* / Q000821:virology; D016022:Case-Control Studies; D018450:Disease Progression; D005260:Female; D017353:Gene Deletion; D006515:Hepatitis B virus / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D019694:Hepatitis B, Chronic / Q000150:complications / Q000821:virology*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods; D006801:Humans; D008113:Liver Neoplasms / Q000453:epidemiology* / Q000821:virology; D008137:Longitudinal Studies; D008297:Male; D008875:Middle Aged; D017354:Point Mutation; D000074704:Quasispecies
keywords: Deep sequencing; Hepatitis B virus; Hepatocellular carcinoma; Pre-S deletions; Quasispecies