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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 2011; 83(7):1121-7JM

Abstract

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.

Authors+Show Affiliations

Influenza Virus Research Center, National Institute of Infectious Diseases, Musashimurayama, Tokyo, Japan.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21567417

Citation

Nakauchi, Mina, et al. "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." Journal of Medical Virology, vol. 83, no. 7, 2011, pp. 1121-7.
Nakauchi M, Ujike M, Obuchi M, et al. 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. 2011;83(7):1121-7.
Nakauchi, M., Ujike, M., Obuchi, M., Takashita, E., Takayama, I., Ejima, M., ... Kageyama, T. (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. Journal of Medical Virology, 83(7), pp. 1121-7. doi:10.1002/jmv.22101.
Nakauchi M, et al. 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. 2011;83(7):1121-7. PubMed PMID: 21567417.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - 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. AU - Nakauchi,Mina, AU - Ujike,Makoto, AU - Obuchi,Masatsugu, AU - Takashita,Emi, AU - Takayama,Ikuyo, AU - Ejima,Miho, AU - Oba,Kunihiro, AU - Konomi,Nami, AU - Odagiri,Takato, AU - Tashiro,Masato, AU - Kageyama,Tsutomu, AU - ,, PY - 2011/5/14/entrez PY - 2011/5/14/pubmed PY - 2011/9/14/medline SP - 1121 EP - 7 JF - Journal of medical virology JO - J. Med. Virol. VL - 83 IS - 7 N2 - 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. SN - 1096-9071 UR - https://www.unboundmedicine.com/medline/citation/21567417/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_ L2 - https://doi.org/10.1002/jmv.22101 DB - PRIME DP - Unbound Medicine ER -