Skip to main content

Advertisement

Log in

HCV inter-subtype 1a/1b recombinant detected by complete-genome next-generation sequencing

  • Original Article
  • Published:
Archives of Virology Aims and scope Submit manuscript

Abstract

Next-generation sequencing (NGS) provides a practical approach to HCV complete-genome sequencing, detecting low-frequency variants and allowing analysis of viral genetic diversity (quasispecies) in the sample, and so far, it is very useful for identifying preexisting drug-resistant mutants and emerging escape mutations, as well as detecting viral recombinants containing genomic regions from different genotypes and subtypes. The aim of this study was to analyze the complete coding region of hepatitis C virus (HCV) genotype 1 (subtypes 1a and 1b) from patients with chronic infection who were direct-acting antiviral (DAA) naïve. Next-generation sequencing (Ion Torrent™ PGM) was used to determine the sequence of the complete coding region of 100 HCV-monoinfected DAA-naïve patients (51 and 49 subtypes 1a and 1b, respectively). We report the first description of nearly complete HCV genome sequences of subtype 1a and 1b isolates from a large population of Brazilian patients with chronic hepatitis C, and HCV-1a grouped in two different clades. Using this methodology, an inter-subtype 1a/1b recombinant was identified in this study.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  1. Mohd Hanafiah K, Groeger J, Flaxman AD, Wiersma ST (2013) Global epidemiology of hepatitis C virus infection: new estimates of age-specific antibody to HCV seroprevalence. Hepatology 57:1333–1342

    Article  PubMed  Google Scholar 

  2. Timm J, Roggendorf M (2007) Sequence diversity of hepatitis C virus: implications for immune control and therapy. World J Gastroenterol 13:4808–4817

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Alavian SM, Ande SR, Coombs KM, Yeganeh B, Davoodpour P, Hashemi M, Los M, Ghavami S (2011) Virus-triggered autophagy in viral hepatitis—possible novel strategies for drug development. J Viral Hepat 18:821–830

    Article  CAS  PubMed  Google Scholar 

  4. Lee LY, Tong CY, Wong T, Wilkinson M (2012) New therapies for chronic hepatitis C infection: a systematic review of evidence from clinical trials. Int J Clin Pract 66:342–355

    Article  CAS  PubMed  Google Scholar 

  5. Yang PL, Gao M, Lin K, Liu Q, Villareal VA (2011) Anti-HCV drugs in the pipeline. Curr Opin Virol 1:607–616

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Smith DB, Bukh J, Kuiken C, Muerhoff AS, Rice CM, Stapleton JT, Simmonds P (2014) Expanded classification of hepatitis C virus into 7 genotypes and 67 subtypes: updated criteria and genotype assignment web resource. Hepatology 59:318–327

    Article  PubMed  Google Scholar 

  7. Luciani F, Bull RA, Lloyd AR (2012) Next generation deep sequencing and vaccine design: today and tomorrow. Trends Biotechnol 30:443–452

    Article  CAS  PubMed  Google Scholar 

  8. Plauzolles A, Lucas M, Gaudieri S (2013) Hepatitis C virus adaptation to T-cell immune pressure. Sci World J 2013:673240

    Article  CAS  Google Scholar 

  9. Xu CH, Shen T, Zheng JJ, Tu J, Zhang WD, Lu FM (2012) Higher dN/dS ratios in the HCV core gene, but not in the E1/HVR1 gene, are associated with human immunodeficiency virus-associated immunosuppression. Arch Virol 157:2153–2162

    Article  CAS  PubMed  Google Scholar 

  10. Morel V, Ghoubra F, Izquierdo L, Martin E, Oliveira C, Francois C, Brochot E, Helle F, Duverlie G, Castelain S (2016) Phylogenetic analysis of a circulating hepatitis C virus recombinant strain 1b/1a isolated in a French hospital centre. Infect Genet Evol 40:374–380

    Article  PubMed  Google Scholar 

  11. Kalinina O, Norder H, Mukomolov S, Magnius LO (2002) A natural intergenotypic recombinant of hepatitis C virus identified in St. Petersburg. J Virol 76:4034–4043

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Kageyama S, Agdamag DM, Alesna ET, Leano PS, Heredia AM, Abellanosa-Tac-An IP, Jereza LD, Tanimoto T, Yamamura J, Ichimura H (2006) A natural inter-genotypic (2b/1b) recombinant of hepatitis C virus in the Philippines. J Med Virol 78:1423–1428

    Article  CAS  PubMed  Google Scholar 

  13. Noppornpanth S, Lien TX, Poovorawan Y, Smits SL, Osterhaus AD, Haagmans BL (2006) Identification of a naturally occurring recombinant genotype 2/6 hepatitis C virus. J Virol 80:7569–7577

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Legrand-Abravanel F, Claudinon J, Nicot F, Dubois M, Chapuy-Regaud S, Sandres-Saune K, Pasquier C, Izopet J (2007) New natural intergenotypic (2/5) recombinant of hepatitis C virus. J Virol 81:4357–4362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Lee YM, Lin HJ, Chen YJ, Lee CM, Wang SF, Chang KY, Chen TL, Liu HF, Chen YM (2010) Molecular epidemiology of HCV genotypes among injection drug users in Taiwan: full-length sequences of two new subtype 6w strains and a recombinant form_2b6w. J Med Virol 82:57–68

    Article  PubMed  Google Scholar 

  16. Colina R, Casane D, Vasquez S, Garcia-Aguirre L, Chunga A, Romero H, Khan B, Cristina J (2004) Evidence of intratypic recombination in natural populations of hepatitis C virus. J Gen Virol 85:31–37

    Article  CAS  PubMed  Google Scholar 

  17. Ross RS, Verbeeck J, Viazov S, Lemey P, Van Ranst M, Roggendorf M (2008) Evidence for a complex mosaic genome pattern in a full-length hepatitis C virus sequence. Evol Bioinform Online 4:249–254

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Moreno P, Alvarez M, Lopez L, Moratorio G, Casane D, Castells M, Castro S, Cristina J, Colina R (2009) Evidence of recombination in hepatitis C virus populations infecting a hemophiliac patient. Virol J 6:203

    Article  PubMed  PubMed Central  Google Scholar 

  19. Calado RA, Rocha MR, Parreira R, Piedade J, Venenno T, Esteves A (2011) Hepatitis C virus subtypes circulating among intravenous drug users in Lisbon, Portugal. J Med Virol 83:608–615

    Article  PubMed  Google Scholar 

  20. Garson JA, Ring C, Tuke P, Tedder RS (1990) Enhanced detection by PCR of hepatitis C virus RNA. Lancet 336:878–879

    Article  CAS  PubMed  Google Scholar 

  21. Alcantara LC, Cassol S, Libin P, Deforche K, Pybus OG, Van Ranst M, Galvao-Castro B, Vandamme AM, de Oliveira T (2009) A standardized framework for accurate, high-throughput genotyping of recombinant and non-recombinant viral sequences. Nucleic Acids Res 37:W634–W642

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Lole KS, Bollinger RC, Paranjape RS, Gadkari D, Kulkarni SS, Novak NG, Ingersoll R, Sheppard HW, Ray SC (1999) Full-length human immunodeficiency virus type 1 genomes from subtype C-infected seroconverters in India, with evidence of intersubtype recombination. J Virol 73:152–160

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Cuypers L, Snoeck J, Vrancken B, Kerremans L, Vuagniaux G, Verbeeck J, Nevens F, Camacho RJ, Vandamme AM, Van Dooren S (2014) A near-full length genotypic assay for HCV1b. J Virol Methods 209:126–135

    Article  CAS  PubMed  Google Scholar 

  24. Capobianchi MR, Giombini E, Rozera G (2013) Next-generation sequencing technology in clinical virology. Clin Microbiol Infect 19:15–22

    Article  CAS  PubMed  Google Scholar 

  25. Peres-da-Silva A, Almeida AJ, Lampe E (2012) Genetic diversity of NS3 protease from Brazilian HCV isolates and possible implications for therapy with direct-acting antiviral drugs. Mem Inst Oswaldo Cruz 107:254–261

    Article  CAS  PubMed  Google Scholar 

  26. Lampe E, Lewis-Ximenez L, Espirito-Santo MP, Delvaux NM, Pereira SA, Peres-da-Silva A, Martins RM, Soares MA, Santos AF, Vidal LL, Germano FN, de Martinez AM, Basso R, Pinho JR, Malta FM, Gomes-Gouvea M, Moliterno RA, Bertolini DA, Fujishima MA, Bello G (2013) Genetic diversity of HCV in Brazil. Antivir Ther 18:435–444

    Article  PubMed  Google Scholar 

  27. Wyles DL, Gutierrez JA (2014) Importance of HCV genotype 1 subtypes for drug resistance and response to therapy. J Viral Hepat 21:229–240

    Article  CAS  PubMed  Google Scholar 

  28. Shi W, Freitas IT, Zhu C, Zheng W, Hall WW, Higgins DG (2012) Recombination in hepatitis C virus: identification of four novel naturally occurring inter-subtype recombinants. PLoS One 7:e41997

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Gonzalez-Candelas F, Lopez-Labrador FX, Bracho MA (2011) Recombination in hepatitis C virus. Viruses 3:2006–2024

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Yun Z, Lara C, Johansson B, Lorenzana de Rivera I, Sonnerborg A (1996) Discrepancy of hepatitis C virus genotypes as determined by phylogenetic analysis of partial NS5 and core sequences. J Med Virol 49:155–160

    Article  CAS  PubMed  Google Scholar 

  31. Du H, Qi Y, Hao F, Huang Y, Mao L, Ji S, Huang M, Qin C, Yan R, Zhu X, Zhang C (2012) Complex patterns of HCV epidemic in Suzhou: evidence for dual infection and HCV recombination in East China. J Clin Virol 54:207–212

    Article  PubMed  Google Scholar 

  32. Hedskog C, Doehle B, Chodavarapu K, Gontcharova V, Crespo Garcia J, De Knegt R, Drenth JP, McHutchison JG, Brainard D, Stamm LM, Miller MD, Svarovskaia E, Mo H (2015) Characterization of hepatitis C virus intergenotypic recombinant strains and associated virological response to sofosbuvir/ribavirin. Hepatology 61:471–480

    Article  CAS  PubMed  Google Scholar 

  33. Lauck M, Alvarado-Mora MV, Becker EA, Bhattacharya D, Striker R, Hughes AL, Carrilho FJ, O’Connor DH, Pinho JR (2012) Analysis of hepatitis C virus intrahost diversity across the coding region by ultradeep pyrosequencing. J Virol 86:3952–3960

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Miura M, Maekawa S, Takano S, Komatsu N, Tatsumi A, Asakawa Y, Shindo K, Amemiya F, Nakayama Y, Inoue T, Sakamoto M, Yamashita A, Moriishi K, Enomoto N (2013) Deep-sequencing analysis of the association between the quasispecies nature of the hepatitis C virus core region and disease progression. J Virol 87:12541–12551

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Akuta N, Suzuki F, Fukushima T, Kawamura Y, Sezaki H, Suzuki Y, Hosaka T, Kobayashi M, Hara T, Kobayashi M, Saitoh S, Arase Y, Ikeda K, Kumada H (2013) Prediction of treatment efficacy and telaprevir-resistant variants after triple therapy in patients infected with hepatitis C virus genotype 1. J Clin Microbiol 51:2862–2868

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Svarovskaia ES, Martin R, McHutchison JG, Miller MD, Mo H (2012) Abundant drug-resistant NS3 mutants detected by deep sequencing in hepatitis C virus-infected patients undergoing NS3 protease inhibitor monotherapy. J Clin Microbiol 50:3267–3274

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Prosperi MC, Yin L, Nolan DJ, Lowe AD, Goodenow MM, Salemi M (2013) Empirical validation of viral quasispecies assembly algorithms: state-of-the-art and challenges. Sci Rep 3:2837

    PubMed  PubMed Central  Google Scholar 

  38. Fan X, Di Bisceglie AM (2010) RT-PCR amplification and cloning of large viral sequences. Methods Mol Biol 630:139–149

    Article  CAS  PubMed  Google Scholar 

  39. Ninomiya M, Ueno Y, Funayama R, Nagashima T, Nishida Y, Kondo Y, Inoue J, Kakazu E, Kimura O, Nakayama K, Shimosegawa T (2011) Use of illumina deep sequencing technology to differentiate hepatitis C virus variants. J Clin Microbiol 50:857–866

    Article  Google Scholar 

Download references

Acknowledgments

Special thanks to the patients, to Dr Maria C. M. Correa for providing samples, and to Dr Ester C. Sabino for allowing the use of next-generation sequencing.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Fernanda de Mello Malta.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Financial support

This project was supported by grant 443152/2014-4 (CNPq) and Alves de Queiroz Family Fund for Research. Karine Vieira Gaspareto is a graduate student from Programa de Pós-Graduação em Biotecnologia, USP, and received a fellowship from CAPES. João Renato Rebello Pinho, Ester Cerdeira Sabino and Maria Cassia Mendes Correa received a fellowship from CNPq (Bolsista de Produtividade).

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Gaspareto, K.V., Ribeiro, R.M., de Mello Malta, F. et al. HCV inter-subtype 1a/1b recombinant detected by complete-genome next-generation sequencing. Arch Virol 161, 2161–2168 (2016). https://doi.org/10.1007/s00705-016-2889-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00705-016-2889-5

Keywords

Navigation