Identification of Oseltamivir Resistance among Pandemic and Seasonal Influenza A (H1N1) Viruses by an His275Tyr Genotyping Assay Using the Cycling Probe Method

Suzuki, Yasushi; Saito, Reiko; Sato, Ichiro; Zaraket, Hassan; Nishikawa, Makoto; Tamura, Tsutomu; Dapat, Clyde; Caperig-Dapat, Isolde; Baranovich, Tatiana; Suzuki, Takahiro; Sano, Hiroshi

doi:10.1128/jcm.01401-10

Cited by 38 publications

(32 citation statements)

References 19 publications

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“…Genotypic analysis of the single-nucleotide polymorphisms M2-S31N and NA-H275Y, which confer resistance to amantadine and oseltamivir, respectively, was performed using cycling probe method [12,13]. In vitro antiviral drug susceptibilities of the isolated viruses were determined by measuring the 50% inhibitory concentrations (IC 50 ) of oseltamivir (Sequoia Research, UK), zanamivir (Sequoia Research, UK), peramivir (Shionogi, Japan), and laninamivir (Daiichi Sankyo, Japan) using a fluorescence-based NA inhibition assay with methylumbelliferone N-acetylneuraminic acid (MUNANA) as the substrate at a final concentration of 0.025 m M [18].…”

Section: Methodsmentioning

confidence: 99%

Characterization of Human Influenza Viruses in Lebanon during 2010-2011 and 2011-2012 Post-Pandemic Seasons

Zaraket¹,

Dapat

Ghanem

et al. 2014

Intervirology

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Objective: To genetically characterize human influenza viruses and their susceptibilities to antivirals during two post-pandemic seasons in Lebanon. Methods: Influenza virus was isolated from nasopharyngeal swabs that were obtained from patients with influenza-like illness during 2010-2012 and further analyzed both phenotypically and genotypically. Results: During the 2010-2011 season, both 2009 pandemic H1N1 (H1N1p) and B viruses co-circulated with equal prevalence, while the H3N2 virus predominated during the 2011-2012 season. All H3N2 and H1N1 viruses were resistant to amantadine. Importantly, all viruses of the influenza A and B types were susceptible to the neuraminidase (NA) inhibitors oseltamivir, zanamivir, peramivir, and laninamivir. Nonetheless, all 2011-2012 H1N1p isolates had three mutations (V241I, N369K, and N386S) in the NA gene that were suggested to be permissive of the H275Y mutation, which confers resistance to oseltamivir. We also detected one H1N1p virus during the 2010-2011 season with a 4-fold decrease in susceptibility to oseltamivir due to an NA-S247N mutation. This isolate was phylogenetically distinct from other H1N1p viruses that were isolated in other regions. Conclusions: Influenza A viruses with reduced susceptibility to oseltamivir and mutations permissive for acquiring NA resistance-conferring mutation with minimal burden on their fitness were isolated in Lebanon.

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Section: Methodsmentioning

confidence: 99%

Characterization of Human Influenza Viruses in Lebanon during 2010-2011 and 2011-2012 Post-Pandemic Seasons

Zaraket¹,

Dapat

Ghanem

et al. 2014

Intervirology

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“…7,8 HA and NA genes were amplified and sequenced as described previously. 9 Phylogenetic tree analyses were performed by using the MEGA program (version 4.0). 10 Reference strains in the tree analyses included some of the closest BLAST search hits for which both the HA and NA genes' sequences were available on the Influenza Virus Resource Data Base (http://www.ncbi.nlm.nih.gov/genomes/FLU/).…”

Section: Virus Isolation and Characterizationmentioning

confidence: 99%

Genetic diversity and antiviral drug resistance of pandemic H1N1 2009 in Lebanon

Zaraket

Kondo

Tabet

et al. 2011

Journal of Clinical Virology

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“…For detection of the NA gene we used a probe labeled with LC610, and for the probe detection the 610 nm channel of LightCycler was used. In previously described tests, the authors used LNA probes [10], High-Resolution Melting [11], the Cycling probe method [18], or two probes for detection of sensitive and resistant strains [13,[19][20][21][22]. There are only a few multiplex assays for detection of influenza virus and determination of oseltamivir sensitivity/resistance, and they are based on conventional PCR [12], or need three probes in the assay [23].…”

Section: Discussionmentioning

confidence: 99%

A multiplex real-time PCR assay for detection of oseltamivir-resistant strains of influenza virus

Nidzworski¹,

Dobkowska²,

Hołysz³

et al. 2014

Open Life Sciences

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Influenza is a contagious disease of humans and animals caused by viruses belonging to the Orthomyxoviridae family. The influenza A virus genome consists of negative sense, single-stranded, segmented RNA. Influenza viruses are classified into subtypes based on two surface antigens known as hemagglutinin (H) and neuraminidase (N). The main problem with influenza A viruses infecting humans is drug resistance, which is caused by antigenic changes. A few antiviral drugs are available, but the most popular is the neuraminidase inhibitor — oseltamivir. The resistance against this drug has probably developed through antigenic drift by a point mutation in one amino acid at position 275 (H275Y). In order to prevent a possible influenza pandemic it is necessary to develop fast diagnostic tests. The aim of this project was to develop a new test for detection of influenza A virus and determination of oseltamivir resistance/sensitivity in humans. Detection and differentiation of oseltamivir resistance/sensitivity of influenza A virus was based on real-time PCR. This test contains two TaqMan probes, which work at different wavelengths. Application of techniques like multiplex real-time PCR has greatly enhanced the capability for surveillance and characterization of influenza viruses. After its potential validation, this test can be used for diagnosis before treatment.

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Identification of Oseltamivir Resistance among Pandemic and Seasonal Influenza A (H1N1) Viruses by an His275Tyr Genotyping Assay Using the Cycling Probe Method

Cited by 38 publications

References 19 publications

Characterization of Human Influenza Viruses in Lebanon during 2010-2011 and 2011-2012 Post-Pandemic Seasons

Characterization of Human Influenza Viruses in Lebanon during 2010-2011 and 2011-2012 Post-Pandemic Seasons

Genetic diversity and antiviral drug resistance of pandemic H1N1 2009 in Lebanon

A multiplex real-time PCR assay for detection of oseltamivir-resistant strains of influenza virus

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