Journal article

A comparison of pyrosequencing and neuraminidase inhibition assays for the detection of oseltamivir-resistant pandemic influenza A(H1N1) 2009 viruses

Yi-Mo Deng, Natalie Caldwell, Aeron Hurt, Tim Shaw, Anne Kelso, Glenys Chidlow, Simon Williams, David Smith, Ian Barr

Antiviral Research | ELSEVIER | Published : 2011


Currently most pandemic influenza A(H1N1) 2009 (H1N1pdm) viruses are sensitive to oseltamivir, but a single point mutation (H275Y) in the neuraminidase (NA) gene of H1N1pdm can lead to resistance and such viruses have been reported from several countries. In this study we compare the performance of a pyrosequencing-based method for the detection of the H275Y mutation in H1N1pdm viruses with a conventional NA inhibition assay. Pyrosequencing could detect as little as 5% H275Y mutants in a mixed viral population, while mixtures with 25% or greater mutant virus were required before a significant increase in IC50 value could be detected. However, the sensitivity of the NA inhibition assay could ..

View full abstract


Awarded by Grants-in-Aid for Scientific Research

Funding Acknowledgements

We thank Dr. Sally Roberts from the Auckland District Health Board, Auckland, New Zealand and Prof. Masato Tashiro from National Institute of Infectious Diseases, Tokyo, Japan for providing viruses used for analysis in this paper. The Melbourne WHO Collaborating Centre for Reference and Research on Influenza is supported by the Australian Government Department of Health and Ageing.