Rapid Detection of Influenza A Pandemic (H1N1) 2009 Virus Neuraminidase Resistance Mutation H275Y by Real-Time Reverse Transcriptase PCR

Hindiyeh, Musa; Ram, Daniela; Mandelboim, Michal; Meningher, Tal; Hirsh, Shira; Robinov, Jana; Levy, Virginia; Orzitzer, Sara; Azar, Roberto; Grossman, Zehava; Mendelson, Ella

doi:10.1128/jcm.02540-09

Cited by 40 publications

(41 citation statements)

References 17 publications

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“…Targeted surveillance directed to the isolation and testing of influenza viruses from immunocompetent or immunocompromised individuals undergoing treatment with NAIs may allow a more focused and thorough assessment of the potential for influenza viruses to develop clinically significant resistance to these compounds. In addition, monitoring could be enhanced by detection of H275Y directly on clinical specimens using molecular methods, including pyrosequencing25, 26, 27 or real‐time RT‐PCR techniques 28, 29…”

Section: Discussionmentioning

confidence: 99%

Neuraminidase inhibitor susceptibility surveillance of influenza viruses circulating worldwide during the 2011 Southern Hemisphere season

Okomo-Adhiambo

Sleeman

Lysen

et al. 2013

Influenza Resp Viruses

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BackgroundNeuraminidase (NA) inhibitors (NAIs) are currently the only antivirals effective against influenza infections due to widespread resistance to M2 inhibitors.MethodsInfluenza A and B viruses (n = 1079) collected worldwide between April 01, 2011, and September 30, 2011, were assessed for susceptibility to FDA‐approved NAIs, oseltamivir and zanamivir, and investigational peramivir, using the fluorescent‐based NA‐Fluor™ Influenza Neuraminidase Assay Kit. A subset of viruses (n = 98) were tested for susceptibility to the investigational NAI, laninamivir.ResultsInfluenza A(H1N1)pdm09 viruses (n = 326) were sensitive to all NAIs, except for two (0·6%) with H275Y (N1 numbering; H274Y in N2 numbering) substitution, which exhibited elevated IC 50s for oseltamivir and peramivir, and a third with previously unreported N325K substitution, exhibiting reduced susceptibility to oseltamivir. Influenza A(H3N2) viruses (n = 407) were sensitive to all NAIs. Influenza B viruses (n = 346) were sensitive to all NAIs, except two (0·6%) with H273Y (N1 numbering; H274Y in N2 numbering) substitution, exhibiting reduced susceptibility to oseltamivir and peramivir, and one with previously unreported G140R and N144K substitutions, exhibiting reduced susceptibility to oseltamivir, zanamivir, and peramivir. All influenza A and B viruses were sensitive to laninamivir. It is unknown whether substitutions N325K, G140R, and N144K were present in the virus prior to culturing because clinical specimens were unavailable for testing.ConclusionsThis study summarizes NAI susceptibility of influenza viruses circulating worldwide during the 2011 Southern Hemisphere (SH) season, assessed using the NA‐Fluor™ Kit. Despite low resistance to NAIs among tested influenza viruses, constant surveillance of influenza virus susceptibility to NAIs should be emphasized.

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Section: Discussionmentioning

confidence: 99%

Neuraminidase inhibitor susceptibility surveillance of influenza viruses circulating worldwide during the 2011 Southern Hemisphere season

Okomo-Adhiambo

Sleeman

Lysen

et al. 2013

Influenza Resp Viruses

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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%

“…There are several molecular methods based on PCR and real-time PCR analysis, but none of them offers a multiplex real-time PCR assay using only two standard TaqMan probes [10][11][12][13].…”

Section: Introductionmentioning

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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“…These include qualitative assays using restriction fragment length polymorphism (18) and qualitative and quantitative real-time reverse transcription (RT)-PCR assay methods (2,5,12,19), some of which can also detect mixtures of wild-type and drug-resistant virus populations.…”

mentioning

confidence: 99%

High-Resolution Melting Approach to Efficient Identification and Quantification of H275Y Mutant Influenza H1N1/2009 Virus in Mixed-Virus-Population Samples

Lee

Loh

et al. 2011

J Clin Microbiol

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Rapid Detection of Influenza A Pandemic (H1N1) 2009 Virus Neuraminidase Resistance Mutation H275Y by Real-Time Reverse Transcriptase PCR

Cited by 40 publications

References 17 publications

Neuraminidase inhibitor susceptibility surveillance of influenza viruses circulating worldwide during the 2011 Southern Hemisphere season

Neuraminidase inhibitor susceptibility surveillance of influenza viruses circulating worldwide during the 2011 Southern Hemisphere season

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

High-Resolution Melting Approach to Efficient Identification and Quantification of H275Y Mutant Influenza H1N1/2009 Virus in Mixed-Virus-Population Samples

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