See supplemental material in the online version of this article for complete list of investigators involved in the influenza virus surveillance group of Japan.
Rapid discrimination of oseltamivir-resistant 275Y and -susceptible 275H substitutions in the neuraminidase gene of pandemic influenza A/H1N1 2009 virus by duplex one-step RT-PCR assay†
Article first published online: 12 MAY 2011
Copyright © 2011 Wiley-Liss, Inc.
Journal of Medical Virology
Volume 83, Issue 7, pages 1121–1127, July 2011
How to Cite
Nakauchi, M., Ujike, M., Obuchi, M., Takashita, E., Takayama, I., Ejima, M., Oba, K., Konomi, N., Odagiri, T., Tashiro, M., Kageyama, T. and the influenza virus surveillance group of Japan (2011), Rapid discrimination of oseltamivir-resistant 275Y and -susceptible 275H substitutions in the neuraminidase gene of pandemic influenza A/H1N1 2009 virus by duplex one-step RT-PCR assay. J. Med. Virol., 83: 1121–1127. doi: 10.1002/jmv.22101
- Issue published online: 12 MAY 2011
- Article first published online: 12 MAY 2011
- Manuscript Accepted: 23 FEB 2011
Errata: Erratum: Rapid discrimination of oseltamivir-resistant 275Y and -susceptible 275H substitutions in the neuraminidase gene of pandemic influenza A/H1N1 2009 virus by duplex one-step RT-PCR assay
Vol. 83, Issue 8, 1419, Article first published online: 15 JUN 2011
- oseltamivir resistant;
- pandemic influenza A/H1N1 2009 virus;
- rapid discrimination
Pandemic influenza A/H1N1 2009 (A/H1N1pdm) virus caused significant outbreaks worldwide last year (2009). A number of oseltamivir-resistant A/H1N1pdm viruses possessing an H275Y substitution in the neuraminidase (NA) protein were reported sporadically in several countries, including Japan, but they were sensitive to zanamivir and did not spread in the community. In this study, to monitor rapidly and simply oseltamivir-resistant A/H1N1pdm viruses possessing H275Y, a duplex one-step RT-PCR assay (H275Y RT-PCR assay) was developed based on an endpoint genotyping analysis method. H275Y RT-PCR assay evaluated using several subtypes/types of influenza A and B viruses and other respiratory pathogenic viruses and shown to have high sensitivity and high specificity. Forty-four clinical specimens were tested after RNA purification using the H275Y RT-PCR assay, resulting in one clinical specimen being found to contain a virus possessing the H275Y mutation. Seventy-three clinical isolates were then tested with the H275Y assay by using clinical isolates in the cultured supernatants of cells directly, without RNA purification, and the results were consistent with the NA sequencing. Since the H275Y RT-PCR assay could detect the H275Y mutation in clinical isolates without RNA purification, as well as a H275Y mutated virus in clinical specimens after RNA purification, the assay was considered a powerful tool for surveillance screening of oseltamivir-resistant A/H1N1pdm virus activity. J. Med. Virol. 83:1121–1127, 2011. © 2011 Wiley-Liss, Inc.