Hepatitis C Virus (HCV) infection remains a significant threat to public health globally. For instance, in sub-Saharan Africa alone, about 10 million people are chronically infected. The virus is genetically diverse and classified into genotypes/sub-genotypes, with varied distribution patterns across the globe. Knowledge of HCV genotype distribution in a given geographic area, as well as virus resistance-associated substitutions, is essential to patient management. Unfortunately, data on the genetic characteristics and diversity of HCV genotypes circulating among low-risk blood donor populations in Kenya are scarce. In this study, we screened for HCV RNA among 450 anti-HCV-positive serum samples collected from voluntary blood donors in Kenya between April 2019 and March 2024 using a real-time RT-qPCR assay. A portion of the virus NSB5 gene was subsequently amplified from three samples that were HCV RNA positive using a hemi-nested PCR and sequenced. Phylogenetic analysis revealed that two of the HCV strains belonged to sub-genotype 1a, while the other belonged to sub-genotype 4v. Mutational analysis indicated that antiviral therapy using sofosbuvir could be effective against these strains. Overall, our results demonstrate the co-circulation of HCV genotypes 1a and 4v among voluntary blood donors in Kenya during the study period. This is the second study to describe the genetic characteristics of HCV among low-risk populations of voluntary blood donors in Kenya. Large-scale prospective genomic surveillance studies on HCV are needed to provide comprehensive insights into virus genotype distribution and resistance-associated substitutions that may impact the efficacy of direct-acting antiviral therapy in infected HCV patients in Kenya.
References
[1]
Hundie, G.B., Raj, V.S., GebreMichael, D., Pas, S.D. and Haagmans, B.L. (2017) Genetic Diversity of Hepatitis C Virus in Ethiopia. PLOSONE, 12, e0179064. https://doi.org/10.1371/journal.pone.0179064
[2]
Garriga, C., Manzanares-Laya, S., García de Olalla, P., Gorrindo, P., Lens, S., Solà, R., et al. (2017) Evolution of Acute Hepatitis C Virus Infection in a Large European City: Trends and New Patterns. PLOSONE, 12, e0187893. https://doi.org/10.1371/journal.pone.0187893
[3]
Cevik, O., Li, D., Baljinnyam, E., Manvar, D., Pimenta, E.M., Waris, G., et al. (2017) Interferon Regulatory Factor 5 (IRF5) Suppresses Hepatitis C Virus (HCV) Replication and HCV-Associated Hepatocellular Carcinoma. JournalofBiologicalChemistry, 292, 21676-21689. https://doi.org/10.1074/jbc.m117.792721
[4]
Farooq, S., Faiz, S., Wahab, A. and Choudhary, M.I. (2024) Determination of Hepatitis C Virus Subtype Prevalent in Sindh, Pakistan: A Phylogenetic Analysis. ScientificReports, 14, Article No. 11159. https://doi.org/10.1038/s41598-024-59342-7
[5]
Petruzziello, A., Marigliano, S., Loquercio, G. and Cacciapuoti, C. (2016) Hepatitis C Virus (HCV) Genotypes Distribution: An Epidemiological Up-Date in Europe. InfectiousAgentsandCancer, 11, Article No. 53. https://doi.org/10.1186/s13027-016-0099-0
[6]
Sonderup, M.W., Afihene, M., Ally, R., Apica, B., Awuku, Y., Cunha, L., et al. (2017) Hepatitis C in Sub-Saharan Africa: The Current Status and Recommendations for Achieving Elimination by 2030. TheLancetGastroenterology&Hepatology, 2, 910-919. https://doi.org/10.1016/s2468-1253(17)30249-2
[7]
Abe, H., Ushijima, Y., Bikangui, R., Ondo, G.N., Pemba, C.M., Zadeh, V.R., et al. (2023) Genetic Diversity of Hepatitis B and C Viruses Revealed by Continuous Surveillance from 2015 to 2021 in Gabon, Central Africa. Microorganisms, 11, Article 2046. https://doi.org/10.3390/microorganisms11082046
[8]
Ashfaq, U.A., Javed, T., Rehman, S., Nawaz, Z. and Riazuddin, S. (2011) An Overview of HCV Molecular Biology, Replication and Immune Responses. VirologyJournal, 8, Article No. 161. https://doi.org/10.1186/1743-422x-8-161
[9]
Nawaz, A., Zaidi, S.F., Usmanghani, K. and Ahmad, I. (2015) Concise Review on the Insight of Hepatitis C. JournalofTaibahUniversityMedicalSciences, 10, 132-139. https://doi.org/10.1016/j.jtumed.2014.08.004
[10]
Dubuisson, J. and Cosset, F. (2014) Virology and Cell Biology of the Hepatitis C Virus Life Cycle—An Update. JournalofHepatology, 61, S3-S13. https://doi.org/10.1016/j.jhep.2014.06.031
[11]
Li, C., Lu, L., Murphy, D.G., Negro, F. and Okamoto, H. (2014) Origin of Hepatitis C Virus Genotype 3 in Africa as Estimated through an Evolutionary Analysis of the Full-Length Genomes of Nine Subtypes, Including the Newly Sequenced 3d and 3e. JournalofGeneralVirology, 95, 1677-1688. https://doi.org/10.1099/vir.0.065128-0
[12]
Gray, R.R., Tanaka, Y., Takebe, Y., Magiorkinis, G., Buskell, Z., Seeff, L., et al. (2013) Evolutionary Analysis of Hepatitis C Virus Gene Sequences from 1953. PhilosophicalTransactionsoftheRoyalSocietyB: BiologicalSciences, 368, Article ID: 20130168. https://doi.org/10.1098/rstb.2013.0168
[13]
Akkarathamrongsin, S., Hacharoen, P., Tangkijvanich, P., Theamboonlers, A., Tanaka, Y., Mizokami, M., et al. (2013) Molecular Epidemiology and Genetic History of Hepatitis C Virus Subtype 3a Infection in Thailand. Intervirology, 56, 284-294. https://doi.org/10.1159/000351621
[14]
Nishiya, A.S., de Almeida-Neto, C., Romano, C.M., Alencar, C.S., Ferreira, S.C., Di-Lorenzo-Oliveira, C., et al. (2015) Phylogenetic Analysis of the Emergence of Main Hepatitis C Virus Subtypes in S?o Paulo, Brazil. TheBrazilianJournalofInfectiousDiseases, 19, 473-478. https://doi.org/10.1016/j.bjid.2015.06.010
[15]
Al-Qahtani, A.A., Baele, G., Khalaf, N., Suchard, M.A., Al-Anazi, M.R., Abdo, A.A., et al. (2017) The Epidemic Dynamics of Hepatitis C Virus Subtypes 4a and 4d in Saudi Arabia. ScientificReports, 7, Article No. 44947. https://doi.org/10.1038/srep44947
[16]
Raghwani, J., Thomas, X.V., Koekkoek, S.M., Schinkel, J., Molenkamp, R., van de Laar, T.J., et al. (2012) Origin and Evolution of the Unique Hepatitis C Virus Circulating Recombinant Form 2k/1b. JournalofVirology, 86, 2212-2220. https://doi.org/10.1128/jvi.06184-11
[17]
Parra, M., Laufer, N., Manrique, J.M., Jones, L.R. and Quarleri, J. (2017) Phylogenetic Diversity in Core Region of Hepatitis C Virus Genotype 1a as a Factor Associated with Fibrosis Severity in HIV-1-Coinfected Patients. BioMedResearchInternational, 2017, Article ID: 1728456. https://doi.org/10.1155/2017/1728456
[18]
Hedskog, C., Chodavarapu, K., Ku, K.S., Xu, S., Martin, R., Miller, M.D., et al. (2015) Genotype-and Subtype-Independent Full-Genome Sequencing Assay for Hepatitis C Virus. JournalofClinicalMicrobiology, 53, 2049-2059. https://doi.org/10.1128/jcm.02624-14
[19]
Levander, S. (2016) Development of Vaccines and Mouse Models for Chronic Hepatitis C Virus Infection. Karolinska Institutet. https://hdl.handle.net/10616/45172
[20]
Candotti, D., Temple, J., Sarkodie, F. and Allain, J. (2003) Frequent Recovery and Broad Genotype 2 Diversity Characterize Hepatitis C Virus Infection in Ghana, West Africa. JournalofVirology, 77, 7914-7923. https://doi.org/10.1128/jvi.77.14.7914-7923.2003
[21]
BioSoft, H. (2014) DNA Sequence Assembler. https://www.dnabaser.com/download/DNA-Baser-sequence-assembler/
[22]
Katoh, K. and Standley, D.M. (2013) MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability. MolecularBiologyandEvolution, 30, 772-780. https://doi.org/10.1093/molbev/mst010
[23]
Nguyen, L., Schmidt, H.A., von Haeseler, A. and Minh, B.Q. (2014) IQ-TREE: A Fast and Effective Stochastic Algorithm for Estimating Maximum-Likelihood Phylogenies. MolecularBiologyandEvolution, 32, 268-274. https://doi.org/10.1093/molbev/msu300
[24]
Rambaut, A. (2010) FigTree v1. 3.1. Institute of Evolutionary Biology, University of Edinburgh. https://tree.bio.ed.ac.uk/software/figtree/
[25]
Onchong’a Robert, M., Okoth Eddy, O., Kimutai Peter, B., Kwallah Allan, O., Gikunda James, M., Ong’ondo Bernard, O., et al. (2020) Characterization of Hepatitis C Virus Circulating among Injecting Drug Users (IDU) in Kilifi County, Kenya. JournalofHumanVirology&Retrovirology, 8, 23-30. https://doi.org/10.15406/jhvrv.2020.08.00217
[26]
Akiyama, M.J., Khudyakov, Y., Ramachandran, S., Riback, L.R., Ackerman, M., Nyakowa, M., et al. (2024) Widespread Hepatitis C Virus Transmission Network among People Who Inject Drugs in Kenya. InternationalJournalofInfectiousDiseases, 147, Article ID: 107215. https://doi.org/10.1016/j.ijid.2024.107215
[27]
Makokha, G.N., Bao, H., Hayes, C.N., Abuduwaili, M., Songok, E., Hijikata, M., et al. (2024) The Prevalence and Genotype Distribution of Hepatitis C Virus in Kenya: A Systematic Review and Meta-Analysis. JournalofEpidemiologyandGlobalHealth, 14, 677-689. https://doi.org/10.1007/s44197-024-00299-1
[28]
Twagirumugabe, T., Swaibu, G., Bergstr?m, T., Walker, T.D., Gahutu, J.B. and Norder, H. (2017) Low Prevalence of Hepatitis C Virus RNA in Blood Donors with Anti-Hepatitis C Virus Reactivity in Rwanda. Transfusion, 57, 2420-2432. https://doi.org/10.1111/trf.14204
[29]
Kamal, S.M. and Nasser, I.A. (2008) Hepatitis C Genotype 4: What We Know and What We Don’t Yet Know. Hepatology, 47, 1371-1383. https://doi.org/10.1002/hep.22127
[30]
Ndong‐Atome, G.R., Makuwa, M., Ouwe‐Missi‐Oukem‐Boyer, O., Pybus, O.G., Branger, M., Le Hello, S., et al. (2008) High Prevalence of Hepatitis C Virus Infection and Predominance of Genotype 4 in Rural Gabon. Journal of Medical Virology, 80, 1581-1587. https://doi.org/10.1002/jmv.21252
[31]
Bulut, M.E. (2020) HCV Genotype Distribution of Patients with Chronic Hepatitis C in Istanbul. SiSliEtfalHastanesiTipBulteni/TheMedicalBulletinofSisliHospital, 55, 86-92. https://doi.org/10.14744/semb.2020.66990
[32]
Bajpai, M., Gupta, E. and Choudhary, A. (2014) Hepatitis C Virus: Screening, Diagnosis, and Interpretation of Laboratory Assays. AsianJournalofTransfusionScience, 8, 19-25. https://doi.org/10.4103/0973-6247.126683
[33]
Schuppan, D., Krebs, A., Bauer, M. and Hahn, E.G. (2003) Hepatitis C and Liver Fibrosis. CellDeath&Differentiation, 10, S59-S67. https://doi.org/10.1038/sj.cdd.4401163
[34]
Silini, E. (1995) Differential Distribution of Hepatitis C Virus Genotypes in Patients with and without Liver Function Abnormalities. Hepatology, 21, 285-290. https://doi.org/10.1016/0270-9139(95)90082-9
[35]
Ahmed, A. and Felmlee, D. (2015) Mechanisms of Hepatitis C Viral Resistance to Direct Acting Antivirals. Viruses, 7, 6716-6729. https://doi.org/10.3390/v7122968
[36]
Tamori, A., Enomoto, M. and Kawada, N. (2016) Recent Advances in Antiviral Therapy for Chronic Hepatitis C. MediatorsofInflammation, 2016, Article ID: 6841628. https://doi.org/10.1155/2016/6841628
[37]
Paolucci, S., Premoli, M., Novati, S., Gulminetti, R., Maserati, R., Barbarini, G., et al. (2017) Baseline and Breakthrough Resistance Mutations in HCV Patients Failing Daas. ScientificReports, 7, Article No. 16017. https://doi.org/10.1038/s41598-017-15987-1