Simultaneous determination of HCV genotype and NS5B resistance associated substitutions using dried serum spots from São Paulo state, Brazil

Kazeem Adeboyejo; Victória Riquena Grosche; Diego Pandeló José; Giulia Magalhães Ferreira; Jacqueline Farinha Shimizu; Barnabas J King; Alexander W Tarr; Márcia Maria Costa Nunes Soares; Jonathan K Ball; C Patrick McClure; Ana Carolina Gomes Jardim

doi:10.1099/acmi.0.000326

Simultaneous determination of HCV genotype and NS5B resistance associated substitutions using dried serum spots from São Paulo state, Brazil

Access Microbiol. 2022 Mar 2;4(3):000326. doi: 10.1099/acmi.0.000326. eCollection 2022.

Authors

Kazeem Adeboyejo^{1

2}, Victória Riquena Grosche^{3

4}, Diego Pandeló José⁵, Giulia Magalhães Ferreira³, Jacqueline Farinha Shimizu^{3

4}, Barnabas J King^{1

2

6}, Alexander W Tarr^{1

2

6}, Márcia Maria Costa Nunes Soares⁷, Jonathan K Ball^{1

2

6}, C Patrick McClure^{1

2

6}, Ana Carolina Gomes Jardim^{3

4}

Affiliations

¹ NIHR Nottingham Biomedical Research Centre, Nottingham University Hospitals NHS Trust and the University of Nottingham, Nottingham, UK.
² School of Life Sciences, University of Nottingham, Nottingham, UK.
³ Institute of Biomedical Sciences, Federal University of Uberlândia, Uberlândia, Minas Gerais, Brazil.
⁴ Institute of Bioscience, Language and Exact Sciences, São Paulo State University, São José do Rio Preto, São Paulo, Brazil.
⁵ Federal University of Triângulo Mineiro, Iturama, Minas Gerais, Brazil.
⁶ MRC/EPSRC Nottingham Molecular Pathology Node, University of Nottingham, Nottingham, UK.
⁷ Instituto Adolfo Lutz, São José do Rio Preto, São Paulo, Brazil.

Abstract

Hepatitis C virus (HCV) is responsible for more than 180 million infections worldwide, and about 80 % of infections are reported in Low and Middle-income countries (LMICs). Therapy is based on the administration of interferon (INF), ribavirin (RBV) or more recently Direct-Acting Antivirals (DAAs). However, amino acid substitutions associated with resistance (RAS) have been extensively described and can contribute to treatment failure, and diagnosis of RAS requires considerable infrastructure, not always locally available. Dried serum spots (DSS) sampling is an alternative specimen collection method, which embeds drops of serum onto filter paper to be transported by posting to a centralized laboratory. Here, we assessed feasibility of genotypic analysis of HCV from DSS in a cohort of 80 patients from São Paulo state Brazil. HCV RNA was detected on DSS specimens in 83 % of samples of HCV infected patients. HCV genotypes 1a, 1b, 2a, 2c and 3a were determined using the sequence of the palm domain of NS5B region, and RAS C316N/Y, Q309R and V321I were identified in HCV 1b samples. Concerning therapy outcome, 75 % of the patients who used INF +RBV as a previous protocol of treatment did not respond to DAAs, and 25 % were end-of-treatment responders. It suggests that therapy with INF plus RBV may contribute for non-response to a second therapeutic protocol with DAAs. One patient that presented RAS (V321I) was classified as non-responder, and combination of RAS C316N and Q309R does not necessarily imply in resistance to treatment in this cohort of patients. Data presented herein highlights the relevance of studying circulating variants for a better understanding of HCV variability and resistance to the therapy. Furthermore, the feasibility of carrying out genotyping and RAS phenotyping analysis by using DSS card for the potential of informing future treatment interventions could be relevant to overcome the limitations of processing samples in several location worldwide, especially in LMICs.

Keywords: Hepatitis C Virus; NS5B; direct-action antivirals; dried serum spot; genotyping; resistance-associated substitutions.

Abstract

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