Hepatitis C virus genotype homogeneity and lack of major Sofosbuvir resistance-associated substitutions among blood donors in Ethiopia

Hepatitis C virus (HCV) is a hepatotropic virus that causes a spectrum of liver diseases. HCV genetic diversity influences transmission, pathogenesis, prognosis, and response to antiviral therapy. Moreover, resistance associated substitutions (RASs) challenged the existing antiviral treatment. However, data regarding circulating genotypes and RASs are limited in Ethiopia. Therefore, this study aimed to determine HCV genotypes and RASs among apparently healthy blood donors. This study used 97 archived anti-HCV-positive serum samples obtained from apparently healthy blood donors, collected in accordance with national routine blood collection practices. The partial HCV NS5B gene of 45 samples was amplified and sequenced using amplicon-based next-generation sequencing. Genotypes and subgenotypes were determined using NCBI BLAST, Geno2Pheno [hcv], and the Los Alamos HCV database, and a phylogenetic tree as a confirmation. The RASs were determined using Geno2Pheno [hcv]. Of the 97 anti-HCV- positive samples, 45 had detectable HCV RNA, of which 35 yielded high-quality sequences. These strains (n = 35) showed marked homogeneity in NS5B gene sequences: 34 were HCV genotype 4, all subgenotype 4d, and one was HCV genotype 2, subgenotype 2c. Most strains carried D310N, an RAS associated with ribavirin, while no RASs associated with sofosbuvir resistance were detected. There is HCV genotype homogeneity with HCV genotype 4, particularly HCV subgenotype 4d, predominating. No major sofosbuvir RASs were found, but ribavirin RAS D310N was common. Future studies should use broader geographic sampling and whole-genome sequencing to comprehensively characterize RASs, their distributions, clinical significance, and temporal trends, thereby guiding treatment strategies in Ethiopia. ImportanceHepatitis C virus infection remains a major global health concern. Its high genetic diversity influences viral evolution, transmission, and response to antiviral therapy. Despite the significant burden of hepatitis C virus infection in sub-Saharan Africa, molecular data on circulating genotypes, subgenotypes, and resistance associated substitutions remain limited. The significance of our research is in characterizing the genetic diversity and resistance associated substitutions among hepatitis C virus strains circulating in Ethiopia. These findings provide important insights into hepatitis C virus molecular epidemiology and may support genomic surveillance, optimization of treatment strategies, and improved understanding of viral evolution trends and patterns among blood donor populations.