Monitoring the susceptibility to oseltamivir of Influenza A(H1N1) 2009 virus by nested-PCR and pyrosequencing during the pandemic and in the season 2010-2011

J Virol Methods. 2012 Sep;184(1-2):113-6. doi: 10.1016/j.jviromet.2012.05.018. Epub 2012 May 29.

Abstract

For the early detection of the H275Y mutation as a marker of oseltamivir resistance in A(H1N1) pandemic strains, a sensitive and specific pyrosequencing assay was developed. This assay analyses a region 99nts long, encompassing the H275Y site, amplified by a nested PCR. Seventy-five respiratory specimens, obtained from 62 patients during the pandemic and in the 2010-2011 influenza season, in Tuscany, were tested. Resistant strains were demonstrated in 10 patients. In three other patients, resistant and sensitive variants were found. This pyrosequencing assay may be a useful method for monitoring the spread of resistant influenza H1N1 2009 strains.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Amino Acid Substitution
  • Antiviral Agents / pharmacology*
  • Child
  • Child, Preschool
  • Humans
  • Infant
  • Infant, Newborn
  • Influenza A Virus, H1N1 Subtype / drug effects*
  • Influenza, Human / virology
  • Italy
  • Microbial Sensitivity Tests / methods
  • Mutation, Missense
  • Oseltamivir / pharmacology*
  • Polymerase Chain Reaction / methods*
  • RNA, Viral / chemistry*
  • RNA, Viral / genetics*
  • Sequence Analysis, DNA / methods*

Substances

  • Antiviral Agents
  • RNA, Viral
  • Oseltamivir