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Article: Resistance to Neuraminidase Inhibitors Conferred by an R292K Mutation in a Human Influenza Virus H7N9 Isolate Can Be Masked by a Mixed R/K Viral Population

TitleResistance to Neuraminidase Inhibitors Conferred by an R292K Mutation in a Human Influenza Virus H7N9 Isolate Can Be Masked by a Mixed R/K Viral Population
Authors
Issue Date2013
PublisherAmerican Society for Microbiology. The Journal's web site is located at http://mbio.asm.org
Citation
mBio, 2013, v. 4 n. 4 How to Cite?
AbstractWe characterized the A/Shanghai/1/2013 virus isolated from the first confirmed human case of A/H7N9 disease in China. The A/Shanghai/1/2013 isolate contained a mixed population of R (65%; 15/23 clones) and K (35%; 8/23 clones) at neuraminidase (NA) residue 292, as determined by clonal sequencing. A/Shanghai/1/2013 with mixed R/K at residue 292 exhibited a phenotype that is sensitive to zanamivir and oseltamivir carboxylate by the enzyme-based NA inhibition assay. The plaque-purified A/Shanghai/1/2013 with dominant K292 (94%; 15/16 clones) showed sensitivity to zanamivir that had decreased by >30-fold and to oseltamivir carboxylate that had decreased by >100-fold compared to its plaque-purified wild-type counterpart possessing dominant R292 (93%, 14/15 clones). In Madin-Darby canine kidney (MDCK) cells, the plaque-purified A/Shanghai/1/2013-NAK292 virus exhibited no reduction in viral titer under conditions of increasing concentrations of oseltamivir carboxylate (range, 0 to 1,000 microM) whereas the replication of the plaque-purified A/Shanghai/1/2013-NAR292 and the A/Shanghai/2/2013 viruses was completely inhibited at 250 microM and 31.25 microM of oseltamivir carboxylate, respectively. Although the plaque-purified A/Shanghai/1/2013-NAK292 virus exhibited lower NA enzyme activity and a higher Km for 2'-(4-methylumbelliferryl)-alpha-d-N-acetylneuraminic acid than the wild-type A/Shanghai/1/2013-NAR292 virus, the A/Shanghai/1/2013-NAK292 virus formed large plaques and replicated efficiently in vitro. Our results confirmed that the NA R292K mutation confers resistance to oseltamivir, peramivir, and zanamivir in the novel human H7N9 viruses. Importantly, detection of the resistance phenotype may be masked in the clinical samples containing a mixed population of R/K at NA residue 292 in the enzyme-based NA inhibition assay. IMPORTANCE: The neuraminidase (NA) inhibitors oseltamivir and zanamivir are currently the front-line therapeutic options against the novel H7N9 influenza viruses, which possess an S31N mutation that confers resistance to the M2 ion channel blockers. It is therefore important to evaluate the sensitivity of the clinical isolates to NA inhibitors and to monitor for the emergence of resistant variants. We characterized the A/Shanghai/1/2013 (H7N9) isolate which contained a mixed population of R/K at NA residue 292. While the clinical isolate exhibited a phenotype of sensitivity to NA inhibitors using the enzyme-based NA inhibition assay, the plaque-purified A/Shanghai/1/2013 virus with dominant K292 was resistant to zanamivir, peramivir, and oseltamivir. Resistance to NA inhibitors conferred by the R292K mutation in a human influenza virus H7N9 isolate can be masked by a mixed R/K viral population, and this should be taken into consideration while monitoring antiviral resistance in patients with H7N9 infection.
Persistent Identifierhttp://hdl.handle.net/10722/188179
ISSN
2015 Impact Factor: 6.975
2015 SCImago Journal Rankings: 3.543
PubMed Central ID
ISI Accession Number ID

 

DC FieldValueLanguage
dc.contributor.authorYen, Hen_US
dc.contributor.authorMcKimm-Breschkin, JLen_US
dc.contributor.authorChoy, KTen_US
dc.contributor.authorWong, DDYen_US
dc.contributor.authorCheung, PPHen_US
dc.contributor.authorZHOU, Jen_US
dc.contributor.authorNg, IHYen_US
dc.contributor.authorZhu, Hen_US
dc.contributor.authorWebby, RJen_US
dc.contributor.authorGuan, Yen_US
dc.contributor.authorWebster, RGen_US
dc.contributor.authorPeiris, JSMen_US
dc.date.accessioned2013-08-21T07:41:47Z-
dc.date.available2013-08-21T07:41:47Z-
dc.date.issued2013en_US
dc.identifier.citationmBio, 2013, v. 4 n. 4en_US
dc.identifier.issn2150-7511-
dc.identifier.urihttp://hdl.handle.net/10722/188179-
dc.description.abstractWe characterized the A/Shanghai/1/2013 virus isolated from the first confirmed human case of A/H7N9 disease in China. The A/Shanghai/1/2013 isolate contained a mixed population of R (65%; 15/23 clones) and K (35%; 8/23 clones) at neuraminidase (NA) residue 292, as determined by clonal sequencing. A/Shanghai/1/2013 with mixed R/K at residue 292 exhibited a phenotype that is sensitive to zanamivir and oseltamivir carboxylate by the enzyme-based NA inhibition assay. The plaque-purified A/Shanghai/1/2013 with dominant K292 (94%; 15/16 clones) showed sensitivity to zanamivir that had decreased by >30-fold and to oseltamivir carboxylate that had decreased by >100-fold compared to its plaque-purified wild-type counterpart possessing dominant R292 (93%, 14/15 clones). In Madin-Darby canine kidney (MDCK) cells, the plaque-purified A/Shanghai/1/2013-NAK292 virus exhibited no reduction in viral titer under conditions of increasing concentrations of oseltamivir carboxylate (range, 0 to 1,000 microM) whereas the replication of the plaque-purified A/Shanghai/1/2013-NAR292 and the A/Shanghai/2/2013 viruses was completely inhibited at 250 microM and 31.25 microM of oseltamivir carboxylate, respectively. Although the plaque-purified A/Shanghai/1/2013-NAK292 virus exhibited lower NA enzyme activity and a higher Km for 2'-(4-methylumbelliferryl)-alpha-d-N-acetylneuraminic acid than the wild-type A/Shanghai/1/2013-NAR292 virus, the A/Shanghai/1/2013-NAK292 virus formed large plaques and replicated efficiently in vitro. Our results confirmed that the NA R292K mutation confers resistance to oseltamivir, peramivir, and zanamivir in the novel human H7N9 viruses. Importantly, detection of the resistance phenotype may be masked in the clinical samples containing a mixed population of R/K at NA residue 292 in the enzyme-based NA inhibition assay. IMPORTANCE: The neuraminidase (NA) inhibitors oseltamivir and zanamivir are currently the front-line therapeutic options against the novel H7N9 influenza viruses, which possess an S31N mutation that confers resistance to the M2 ion channel blockers. It is therefore important to evaluate the sensitivity of the clinical isolates to NA inhibitors and to monitor for the emergence of resistant variants. We characterized the A/Shanghai/1/2013 (H7N9) isolate which contained a mixed population of R/K at NA residue 292. While the clinical isolate exhibited a phenotype of sensitivity to NA inhibitors using the enzyme-based NA inhibition assay, the plaque-purified A/Shanghai/1/2013 virus with dominant K292 was resistant to zanamivir, peramivir, and oseltamivir. Resistance to NA inhibitors conferred by the R292K mutation in a human influenza virus H7N9 isolate can be masked by a mixed R/K viral population, and this should be taken into consideration while monitoring antiviral resistance in patients with H7N9 infection.-
dc.languageengen_US
dc.publisherAmerican Society for Microbiology. The Journal's web site is located at http://mbio.asm.org-
dc.relation.ispartofmBioen_US
dc.rightsmBio. Copyright © American Society for Microbiology.-
dc.rightsCopyright © American Society for Microbiology, [mBio, v. 4, 2013]-
dc.subject.meshCoinfection - virology-
dc.subject.meshDrug Resistance, Viral-
dc.subject.meshInfluenza A Virus, H7N9 Subtype - drug effects - genetics - isolation and purification-
dc.subject.meshInfluenza, Human - virology-
dc.subject.meshMutation, Missense-
dc.titleResistance to Neuraminidase Inhibitors Conferred by an R292K Mutation in a Human Influenza Virus H7N9 Isolate Can Be Masked by a Mixed R/K Viral Populationen_US
dc.typeArticleen_US
dc.identifier.emailYen, H: hyen@hku.hken_US
dc.identifier.emailChoy, KT: ktchoy@hku.hken_US
dc.identifier.emailWong, DDY: dywong@hku.hken_US
dc.identifier.emailCheung, PPH: pcheun5@hku.hken_US
dc.identifier.emailNg, IHY: ihyng@hkucc.hku.hken_US
dc.identifier.emailZhu, H: zhuhch@hku.hken_US
dc.identifier.emailGuan, Y: yguan@hkucc.hku.hken_US
dc.identifier.emailPeiris, JSM: malik@hkucc.hku.hken_US
dc.identifier.authorityYen, H=rp00304en_US
dc.identifier.authorityZhu, H=rp01535en_US
dc.identifier.authorityGuan, Y=rp00397en_US
dc.identifier.authorityPeiris, JSM=rp00410en_US
dc.description.naturelink_to_OA_fulltext-
dc.identifier.doi10.1128/mBio.00396-13-
dc.identifier.pmid23860768-
dc.identifier.pmcidPMC3735122-
dc.identifier.hkuros217326en_US
dc.identifier.volume4-
dc.identifier.issue4-
dc.identifier.isiWOS:000326881100026-
dc.publisher.placeUnited States-

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