AIM To evaluate the occurrence of resistant mutations in treatment-na?ve hepatitis C virus(HCV) sequences deposited in the European hepatitis C virus database(euH CVdb). METHODS The sequences were downloaded from the ...AIM To evaluate the occurrence of resistant mutations in treatment-na?ve hepatitis C virus(HCV) sequences deposited in the European hepatitis C virus database(euH CVdb). METHODS The sequences were downloaded from the eu HCVdb(https://euhcvdb.ibcp.fr/eu HCVdb/). The search was performed for full-length NS3 protease, NS5 A and NS5 B polymerase sequences of HCV, separated by genotypes 1a, 1b, 2a, 2b and 3a, and resulted in 798 NS3, 708 NS5 A and 535 NS5 B sequences from HCV genotypes1a, 1b, 2a, 2b and 3a, after the exclusion of sequences containing errors and/or gaps or incomplete sequences, and sequences from patients previously treated with direct antiviral agents(DAA). The sequence alignment was performed with MEGA 6.06 MAC and the resulting protein sequences were then analyzed using the BioE dit 7.2.5. for mutations associated with resistance. Only positions that have been described as being associated with failure in treatment in in vivo studies, and/or as conferring a more than 2-fold change in replication in comparison to the wildtype reference strain in in vitro phenotypic assays were included in the analysis.RESULTS The Q80 K variant in the NS3 gene was the most prevalent mutation, being found in 44.66% of subtype 1a and 0.25% of subtype 1b. Other frequent mutations observed in more than 2% of the NS3 sequences were: I170V(3.21%) in genotype 1a, and Y56F(15.93%), V132I(23.28%) and I170V(65.20%) in genotype 1b. For the NS5 A, 2.21% of the genotype 1a sequences have the P58 S mutation, 5.95% of genotype 1b sequences have the R30 Q mutation, 15.79% of subtypes 2a sequences have the Q30 R mutation, 23.08% of subtype 2b sequences have a L31 M mutation, and in subtype 3a sequences, 23.08% have the M31 L resistant variants. For the NS5 B, the V321 L RAV was identified in 0.60% of genotype 1a and in 0.32% of genotype 1b sequences, and the N142 T variant was observed in 0.32% of subtype 1b sequences. The C316 Y, S556 G, D559 N RAV were identified in 0.33%, 7.82% and 0.32% of genotype 1b sequences, respectively, and were not observed in other genotypes.CONCLUSION HCV mutants resistant to DAAs are found in low frequency, nevertheless they could be selected and therapy could fail due resistance substitutions in HCV genome.展开更多
文摘AIM To evaluate the occurrence of resistant mutations in treatment-na?ve hepatitis C virus(HCV) sequences deposited in the European hepatitis C virus database(euH CVdb). METHODS The sequences were downloaded from the eu HCVdb(https://euhcvdb.ibcp.fr/eu HCVdb/). The search was performed for full-length NS3 protease, NS5 A and NS5 B polymerase sequences of HCV, separated by genotypes 1a, 1b, 2a, 2b and 3a, and resulted in 798 NS3, 708 NS5 A and 535 NS5 B sequences from HCV genotypes1a, 1b, 2a, 2b and 3a, after the exclusion of sequences containing errors and/or gaps or incomplete sequences, and sequences from patients previously treated with direct antiviral agents(DAA). The sequence alignment was performed with MEGA 6.06 MAC and the resulting protein sequences were then analyzed using the BioE dit 7.2.5. for mutations associated with resistance. Only positions that have been described as being associated with failure in treatment in in vivo studies, and/or as conferring a more than 2-fold change in replication in comparison to the wildtype reference strain in in vitro phenotypic assays were included in the analysis.RESULTS The Q80 K variant in the NS3 gene was the most prevalent mutation, being found in 44.66% of subtype 1a and 0.25% of subtype 1b. Other frequent mutations observed in more than 2% of the NS3 sequences were: I170V(3.21%) in genotype 1a, and Y56F(15.93%), V132I(23.28%) and I170V(65.20%) in genotype 1b. For the NS5 A, 2.21% of the genotype 1a sequences have the P58 S mutation, 5.95% of genotype 1b sequences have the R30 Q mutation, 15.79% of subtypes 2a sequences have the Q30 R mutation, 23.08% of subtype 2b sequences have a L31 M mutation, and in subtype 3a sequences, 23.08% have the M31 L resistant variants. For the NS5 B, the V321 L RAV was identified in 0.60% of genotype 1a and in 0.32% of genotype 1b sequences, and the N142 T variant was observed in 0.32% of subtype 1b sequences. The C316 Y, S556 G, D559 N RAV were identified in 0.33%, 7.82% and 0.32% of genotype 1b sequences, respectively, and were not observed in other genotypes.CONCLUSION HCV mutants resistant to DAAs are found in low frequency, nevertheless they could be selected and therapy could fail due resistance substitutions in HCV genome.
