Detection of H275Y oseltamivir resistance gene mutation among Influenza A(H1N1)pdm09 patients by allelic discrimination real-time RT-PCR

J Med Virol. 2023 May;95(5):e28764. doi: 10.1002/jmv.28764.

Abstract

Influenza viruses can mutate genetically and cause a range of respiratory ailments. The H275Y mutation in the neuraminidase (NA) gene reduces the effectiveness of oseltamivir, a widely used drug for the treatment of Influenza A and B virus infection. The World Health Organization (WHO) recommends single-nucleotide polymorphism assays to detect this mutation. The present study aims to estimate the prevalence of H275Y mutation conferring oseltamivir resistance in Influenza A(H1N1)pdm09 virus among hospitalized patients from June 2014 to December 2021. Following the WHO protocol, allelic discrimination real-time RT-PCR was performed for 752 samples. Out of the 752 samples, 1 tested positive for Y275 gene mutation by allelic discrimination real-time RT-PCR. In samples of years 2020 and 2021, neither the H275 nor Y275 genotype was detected. Sequencing of the NA gene of all negative samples showed a mismatch between the NA sequence and the probes used in the allelic discrimination assay. Also, Y275 mutation was detected in only 1 sample from 2020. The prevalence of oseltamivir resistance was estimated as 0.27% among the Influenza A(H1N1)pdm09 patients during 2014-2021. The study highlights that the WHO-recommended probes for detecting H275Y mutation may not be useful to detect 2020 and 2021 circulating strains of Influenza A(H1N1)pdm09, emphasizing the need for continuous monitoring of mutations in the influenza virus.

Keywords: H275Y; Influenza A(H1N1)pdm09 virus; allelic discrimination real-time RT-PCR; oseltamivir resistance.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antiviral Agents / pharmacology
  • Antiviral Agents / therapeutic use
  • Drug Resistance, Viral / genetics
  • Humans
  • Influenza A Virus, H1N1 Subtype* / genetics
  • Influenza A virus* / genetics
  • Influenza, Human* / drug therapy
  • Influenza, Human* / epidemiology
  • Mutation
  • Mutation, Missense
  • Neuraminidase / genetics
  • Oseltamivir / pharmacology
  • Oseltamivir / therapeutic use
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Oseltamivir
  • Antiviral Agents
  • Neuraminidase