An NS5A single optimized method to determine genotype, subtype and resistance profiles of Hepatitis C strains

Abstract : The objective was to develop a method of HCV genome sequencing that allowed simultaneous genotyping and NS5A inhibitor resistance profiling. In order to validate the use of a unique RT-PCR for genotypes 1-5, 142 plasma samples from patients infected with HCV were analysed. The NS4B-NS5A partial region was successfully amplified and sequenced in all samples. In parallel, partial NS3 sequences were analyzed obtained for genotyping. Phylogenetic analysis showed concordance of genotypes and subtypes with a bootstrap >95% for each type cluster. NS5A resistance mutations were analyzed using the Geno2-pheno [hcv] v0.92 tool and compared to the list of known Resistant Associated Substitutions recently published. In conclusion, this tool allows determination of HCV genotypes, sub-types and identification of NS5A resistance mutations. This single method can be used to detect pre-existing resistance mutations in NS5A before treatment and to check the emergence of resistant viruses while undergoing treatment in major HCV genotypes (G1-5) in the EU and the US.
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Elisabeth André-Garnier, Bernard Besse, Audrey Rodallec, Olivier Ribeyrol, Virginie Ferre, et al.. An NS5A single optimized method to determine genotype, subtype and resistance profiles of Hepatitis C strains. PLoS ONE, Public Library of Science, 2017. ⟨inserm-02155275⟩

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