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Published in: Virology Journal 1/2019

Open Access 01-12-2019 | Hepatitis C | Research

Hepatitis C virus genotyping based on Core and NS5B regions in Cameroonian patients

Authors: Paul Alain Tagnouokam-Ngoupo, Marie Nicole Ngoufack, Sebastien Kenmoe, Simon Frédéric Lissock, Marie Amougou-Atsama, Robert Banai, Laure Ngono, Richard Njouom

Published in: Virology Journal | Issue 1/2019

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Abstract

Background

Current HCV treatments are genotype specific although potential pan-genotype treatments have recently been described. Therefore, genotyping is an essential tool for the therapeutic management of HCV infection and a variety of technologies have been developed for HCV genotypes determination. Sequences analysis of HCV sub-genomic regions is considered as gold standard and is widely used for HCV genotyping. Here, we compared HCV genotyping using core and NS5B regions in routine practice in HCV-positive Cameroonian patients.

Methods

All plasma samples received at Centre Pasteur of Cameroon (CPC) in 2016 for HCV genotyping were included. Viral loads were determined using the Abbott Real Time assay. Further, genotyping was based on the amplification and sequencing of core and NS5B regions following by phylogenetic analysis of corresponding sequences.

Results

A total of 369 samples were received during the study period with high viral load values (median: 930,952 IU/ml; IQR: 281,833-2,861,179). Positive amplification was obtained in at least one genomic region (core or NS5B) for all the samples with similar amplification rate in the two genomic regions (p = 0.34). Phylogenetic analysis showed that among the 369 samples, 146 (39.6%) were classified as genotype 4, 132 (35.8%) as genotype 1, 89 (24.1%) as genotype 2, in both core and NS5B regions. Interestingly, for two samples (0.54%) discordant genotypes were obtained in both regions with the core region classified as genotype 4 while the NS5B was identified as genotype 1 indicating the presence of putative HCV recombinant virus or multiple infections in these samples. Discrimination of HCV subtypes was most likely possible with NS5B compared to core region.

Conclusions

We found high amplification rates of HCV in both core and NS5B regions, and a good concordance was obtained at genotype level using both regions except for two samples where putative 1–4 recombinants/multiple infections were detected. Therefore, HCV genotyping based on at least two genomic regions could help to identify putative recombinants and improve therapeutic management of HCV infection.
Literature
1.
go back to reference Petruzziello A, Marigliano S, Loquercio G, Cozzolino A, Cacciapuoti C. Global epidemiology of hepatitis C virus infection: an up-date of the distribution and circulation of hepatitis C virus genotypes. World J Gastroenterol. 2016;22:7824–40.CrossRef Petruzziello A, Marigliano S, Loquercio G, Cozzolino A, Cacciapuoti C. Global epidemiology of hepatitis C virus infection: an up-date of the distribution and circulation of hepatitis C virus genotypes. World J Gastroenterol. 2016;22:7824–40.CrossRef
2.
go back to reference Smith DB, Bukh J, Kuiken C, Muerhoff AS, Rice CM, Stapleton JT, Simmonds P. Expanded classification of hepatitis C virus into 7 genotypes and 67 subtypes: updated criteria and genotype assignment web resource. Hepatology. 2014;59:318–27.CrossRef Smith DB, Bukh J, Kuiken C, Muerhoff AS, Rice CM, Stapleton JT, Simmonds P. Expanded classification of hepatitis C virus into 7 genotypes and 67 subtypes: updated criteria and genotype assignment web resource. Hepatology. 2014;59:318–27.CrossRef
3.
go back to reference Pasquier C, Njouom R, Ayouba A, Dubois M, Sartre MT, Vessiere A, Timba I, Thonnon J, Izopet J, Nerrienet E. Distribution and heterogeneity of hepatitis C genotypes in hepatitis patients in Cameroon. J Med Virol. 2005;77:390–8.CrossRef Pasquier C, Njouom R, Ayouba A, Dubois M, Sartre MT, Vessiere A, Timba I, Thonnon J, Izopet J, Nerrienet E. Distribution and heterogeneity of hepatitis C genotypes in hepatitis patients in Cameroon. J Med Virol. 2005;77:390–8.CrossRef
4.
go back to reference Njouom R, Pasquier C, Ayouba A, Sandres-Saune K, Mfoupouendoun J, Mony Lobe M, Tene G, Thonnon J, Izopet J, Nerrienet E. Hepatitis C virus infection among pregnant women in Yaounde, Cameroon: prevalence, viremia, and genotypes. J Med Virol. 2003;69:384–90.CrossRef Njouom R, Pasquier C, Ayouba A, Sandres-Saune K, Mfoupouendoun J, Mony Lobe M, Tene G, Thonnon J, Izopet J, Nerrienet E. Hepatitis C virus infection among pregnant women in Yaounde, Cameroon: prevalence, viremia, and genotypes. J Med Virol. 2003;69:384–90.CrossRef
5.
go back to reference D'Ambrosio R, Degasperi E, Colombo M, Aghemo A. Direct-acting antivirals: the endgame for hepatitis C? Curr Opin Virol. 2017;24:31–7.CrossRef D'Ambrosio R, Degasperi E, Colombo M, Aghemo A. Direct-acting antivirals: the endgame for hepatitis C? Curr Opin Virol. 2017;24:31–7.CrossRef
6.
go back to reference Tong L, Yu W, Chen L, Selyutin O, Dwyer MP, Nair AG, Mazzola R, Kim JH, Sha D, Yin J, et al. Discovery of Ruzasvir (MK-8408): a potent, pan-genotype HCV NS5A inhibitor with optimized activity against common resistance-associated polymorphisms. J Med Chem. 2017;60:290–306.CrossRef Tong L, Yu W, Chen L, Selyutin O, Dwyer MP, Nair AG, Mazzola R, Kim JH, Sha D, Yin J, et al. Discovery of Ruzasvir (MK-8408): a potent, pan-genotype HCV NS5A inhibitor with optimized activity against common resistance-associated polymorphisms. J Med Chem. 2017;60:290–306.CrossRef
7.
go back to reference Yu W, Coburn CA, Yang DY, Meinke PT, Wong M, Rosenblum SB, Chen KX, Njoroge GF, Chen L, Dwyer MP, et al. Discovery of fused tricyclic core containing HCV NS5A inhibitors with pan-genotype activity. Bioorg Med Chem Lett. 2016;26:3158–62.CrossRef Yu W, Coburn CA, Yang DY, Meinke PT, Wong M, Rosenblum SB, Chen KX, Njoroge GF, Chen L, Dwyer MP, et al. Discovery of fused tricyclic core containing HCV NS5A inhibitors with pan-genotype activity. Bioorg Med Chem Lett. 2016;26:3158–62.CrossRef
8.
go back to reference WHO: Guidelines for the care and treatment of persons diagnosed with chronic hepatitis c virus infection. 2018. WHO: Guidelines for the care and treatment of persons diagnosed with chronic hepatitis c virus infection. 2018.
9.
go back to reference Maughan A, Sadigh K, Angulo-Diaz V, Mandimika C, Villanueva M, Lim JK, Ogbuagu O. Contemporary HCV pangenotypic DAA treatment protocols are exclusionary to real world HIV-HCV co-infected patients. BMC Infect Dis. 2019;19:378.CrossRef Maughan A, Sadigh K, Angulo-Diaz V, Mandimika C, Villanueva M, Lim JK, Ogbuagu O. Contemporary HCV pangenotypic DAA treatment protocols are exclusionary to real world HIV-HCV co-infected patients. BMC Infect Dis. 2019;19:378.CrossRef
10.
go back to reference Buoro S, Pizzighella S, Boschetto R, Pellizzari L, Cusan M, Bonaguro R, Mengoli C, Caudai C, Padula M, Egisto Valensin P, Palu G. Typing of hepatitis C virus by a new method based on restriction fragment length polymorphism. Intervirology. 1999;42:1–8.CrossRef Buoro S, Pizzighella S, Boschetto R, Pellizzari L, Cusan M, Bonaguro R, Mengoli C, Caudai C, Padula M, Egisto Valensin P, Palu G. Typing of hepatitis C virus by a new method based on restriction fragment length polymorphism. Intervirology. 1999;42:1–8.CrossRef
11.
go back to reference Stuyver L, Rossau R, Wyseur A, Duhamel M, Vanderborght B, Van Heuverswyn H, Maertens G: Typing of hepatitis C virus isolates and characterization of new subtypes using a line probe assay. J Gen Virol 1993;74 ( Pt 6):1093–1102.CrossRef Stuyver L, Rossau R, Wyseur A, Duhamel M, Vanderborght B, Van Heuverswyn H, Maertens G: Typing of hepatitis C virus isolates and characterization of new subtypes using a line probe assay. J Gen Virol 1993;74 ( Pt 6):1093–1102.CrossRef
12.
go back to reference Avo AP, Agua-Doce I, Andrade A, Padua E. Hepatitis C virus subtyping based on sequencing of the C/E1 and NS5B genomic regions in comparison to a commercially available line probe assay. J Med Virol. 2013;85:815–22.CrossRef Avo AP, Agua-Doce I, Andrade A, Padua E. Hepatitis C virus subtyping based on sequencing of the C/E1 and NS5B genomic regions in comparison to a commercially available line probe assay. J Med Virol. 2013;85:815–22.CrossRef
13.
go back to reference Nakatani SM, Santos CA, Riediger IN, Krieger MA, Duarte CA, do Carmo Debur M, Carrilho FJ, Ono SK. Comparative performance evaluation of hepatitis C virus genotyping based on the 5′ untranslated region versus partial sequencing of the NS5B region of brazilian patients with chronic hepatitis C. Virol J. 2011;8:459.CrossRef Nakatani SM, Santos CA, Riediger IN, Krieger MA, Duarte CA, do Carmo Debur M, Carrilho FJ, Ono SK. Comparative performance evaluation of hepatitis C virus genotyping based on the 5′ untranslated region versus partial sequencing of the NS5B region of brazilian patients with chronic hepatitis C. Virol J. 2011;8:459.CrossRef
14.
go back to reference Simmonds P, Smith DB, McOmish F, Yap PL, Kolberg J, Urdea MS, Holmes EC: Identification of genotypes of hepatitis C virus by sequence comparisons in the core, E1 and NS-5 regions. J Gen Virol 1994;75 ( Pt 5):1053–1061.CrossRef Simmonds P, Smith DB, McOmish F, Yap PL, Kolberg J, Urdea MS, Holmes EC: Identification of genotypes of hepatitis C virus by sequence comparisons in the core, E1 and NS-5 regions. J Gen Virol 1994;75 ( Pt 5):1053–1061.CrossRef
15.
go back to reference Cai Q, Zhao Z, Liu Y, Shao X, Gao Z. Comparison of three different HCV genotyping methods: core, NS5B sequence analysis and line probe assay. Int J Mol Med. 2013;31:347–52.CrossRef Cai Q, Zhao Z, Liu Y, Shao X, Gao Z. Comparison of three different HCV genotyping methods: core, NS5B sequence analysis and line probe assay. Int J Mol Med. 2013;31:347–52.CrossRef
16.
go back to reference Kassela K, Karakasiliotis I, Kokkiou E, Souvalidou F, Mimidis P, Veletza S, Panopoulou M, Koskinas J, Mimidis K, Mavromara P. Intergenotypic 2k/1b hepatitis C virus recombinants in the East Macedonia and Thrace region of Greece. Ann Gastroenterol. 2019;32:88–92.PubMed Kassela K, Karakasiliotis I, Kokkiou E, Souvalidou F, Mimidis P, Veletza S, Panopoulou M, Koskinas J, Mimidis K, Mavromara P. Intergenotypic 2k/1b hepatitis C virus recombinants in the East Macedonia and Thrace region of Greece. Ann Gastroenterol. 2019;32:88–92.PubMed
17.
go back to reference Zakalashvili M, Zarkua J, Weizenegger M, Bartel J, Raabe M, Zangurashvili L, Kankia N, Jashiashvili N, Lomidze M, Telia T, et al. Identification of hepatitis C virus 2k/1b intergenotypic recombinants in Georgia. Liver Int. 2018;38:451–7.CrossRef Zakalashvili M, Zarkua J, Weizenegger M, Bartel J, Raabe M, Zangurashvili L, Kankia N, Jashiashvili N, Lomidze M, Telia T, et al. Identification of hepatitis C virus 2k/1b intergenotypic recombinants in Georgia. Liver Int. 2018;38:451–7.CrossRef
18.
go back to reference Iles JC, Njouom R, Foupouapouognigni Y, Bonsall D, Bowden R, Trebes A, Piazza P, Barnes E, Pepin J, Klenerman P, Pybus OG. Characterization of hepatitis C virus recombination in Cameroon by use of nonspecific next-generation sequencing. J Clin Microbiol. 2015;53:3155–64.CrossRef Iles JC, Njouom R, Foupouapouognigni Y, Bonsall D, Bowden R, Trebes A, Piazza P, Barnes E, Pepin J, Klenerman P, Pybus OG. Characterization of hepatitis C virus recombination in Cameroon by use of nonspecific next-generation sequencing. J Clin Microbiol. 2015;53:3155–64.CrossRef
19.
go back to reference Kalinina O, Norder H, Mukomolov S, Magnius LO. A natural intergenotypic recombinant of hepatitis C virus identified in St. Petersburg. J Virol. 2002;76:4034–43.CrossRef Kalinina O, Norder H, Mukomolov S, Magnius LO. A natural intergenotypic recombinant of hepatitis C virus identified in St. Petersburg. J Virol. 2002;76:4034–43.CrossRef
20.
go back to reference Legrand-Abravanel F, Claudinon J, Nicot F, Dubois M, Chapuy-Regaud S, Sandres-Saune K, Pasquier C, Izopet J. New natural intergenotypic (2/5) recombinant of hepatitis C virus. J Virol. 2007;81:4357–62.CrossRef Legrand-Abravanel F, Claudinon J, Nicot F, Dubois M, Chapuy-Regaud S, Sandres-Saune K, Pasquier C, Izopet J. New natural intergenotypic (2/5) recombinant of hepatitis C virus. J Virol. 2007;81:4357–62.CrossRef
21.
go back to reference Morel V, Fournier C, Francois C, Brochot E, Helle F, Duverlie G, Castelain S. Genetic recombination of the hepatitis C virus: clinical implications. J Viral Hepat. 2011;18:77–83.CrossRef Morel V, Fournier C, Francois C, Brochot E, Helle F, Duverlie G, Castelain S. Genetic recombination of the hepatitis C virus: clinical implications. J Viral Hepat. 2011;18:77–83.CrossRef
22.
go back to reference Gonzalez-Candelas F, Lopez-Labrador FX, Bracho MA. Recombination in hepatitis C virus. Viruses. 2011;3:2006–24.CrossRef Gonzalez-Candelas F, Lopez-Labrador FX, Bracho MA. Recombination in hepatitis C virus. Viruses. 2011;3:2006–24.CrossRef
23.
go back to reference Lole KS, Jha JA, Shrotri SP, Tandon BN, Prasad VG, Arankalle VA. Comparison of hepatitis C virus genotyping by 5′ noncoding region- and core-based reverse transcriptase PCR assay with sequencing and use of the assay for determining subtype distribution in India. J Clin Microbiol. 2003;41:5240–4.CrossRef Lole KS, Jha JA, Shrotri SP, Tandon BN, Prasad VG, Arankalle VA. Comparison of hepatitis C virus genotyping by 5′ noncoding region- and core-based reverse transcriptase PCR assay with sequencing and use of the assay for determining subtype distribution in India. J Clin Microbiol. 2003;41:5240–4.CrossRef
24.
go back to reference Njouom R, Caron M, Besson G, Ndong-Atome GR, Makuwa M, Pouillot R, Nkoghe D, Leroy E, Kazanji M. Phylogeography, risk factors and genetic history of hepatitis C virus in Gabon, Central Africa. PLoS One. 2012;7:e42002.CrossRef Njouom R, Caron M, Besson G, Ndong-Atome GR, Makuwa M, Pouillot R, Nkoghe D, Leroy E, Kazanji M. Phylogeography, risk factors and genetic history of hepatitis C virus in Gabon, Central Africa. PLoS One. 2012;7:e42002.CrossRef
25.
go back to reference Laperche S, Lunel F, Izopet J, Alain S, Deny P, Duverlie G, Gaudy C, Pawlotsky JM, Plantier JC, Pozzetto B, et al. Comparison of hepatitis C virus NS5b and 5′ noncoding gene sequencing methods in a multicenter study. J Clin Microbiol. 2005;43:733–9.CrossRef Laperche S, Lunel F, Izopet J, Alain S, Deny P, Duverlie G, Gaudy C, Pawlotsky JM, Plantier JC, Pozzetto B, et al. Comparison of hepatitis C virus NS5b and 5′ noncoding gene sequencing methods in a multicenter study. J Clin Microbiol. 2005;43:733–9.CrossRef
26.
go back to reference Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol. 2013;30:2725–9.CrossRef Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol. 2013;30:2725–9.CrossRef
27.
go back to reference Ngoupo PA, Sadeuh-Mba SA, De Oliveira F, Ngono V, Ngono L, Tchendjou P, Penlap V, Mourez T, Njouom R, Kfutwah A, Plantier JC. First evidence of transmission of an HIV-1 M/O intergroup recombinant virus. Aids. 2016;30:1–8.CrossRef Ngoupo PA, Sadeuh-Mba SA, De Oliveira F, Ngono V, Ngono L, Tchendjou P, Penlap V, Mourez T, Njouom R, Kfutwah A, Plantier JC. First evidence of transmission of an HIV-1 M/O intergroup recombinant virus. Aids. 2016;30:1–8.CrossRef
28.
go back to reference Murphy DG, Willems B, Deschenes M, Hilzenrat N, Mousseau R, Sabbah S. Use of sequence analysis of the NS5B region for routine genotyping of hepatitis C virus with reference to C/E1 and 5′ untranslated region sequences. J Clin Microbiol. 2007;45:1102–12.CrossRef Murphy DG, Willems B, Deschenes M, Hilzenrat N, Mousseau R, Sabbah S. Use of sequence analysis of the NS5B region for routine genotyping of hepatitis C virus with reference to C/E1 and 5′ untranslated region sequences. J Clin Microbiol. 2007;45:1102–12.CrossRef
Metadata
Title
Hepatitis C virus genotyping based on Core and NS5B regions in Cameroonian patients
Authors
Paul Alain Tagnouokam-Ngoupo
Marie Nicole Ngoufack
Sebastien Kenmoe
Simon Frédéric Lissock
Marie Amougou-Atsama
Robert Banai
Laure Ngono
Richard Njouom
Publication date
01-12-2019
Publisher
BioMed Central
Keyword
Hepatitis C
Published in
Virology Journal / Issue 1/2019
Electronic ISSN: 1743-422X
DOI
https://doi.org/10.1186/s12985-019-1214-9

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