title: Pyrosequencing reveals an oseltamivir-resistant marker in the quasispecies of avian influenza A (H7N9) virus.
abstract: Prompt diagnosis of an oseltamivir-resistant marker is important for patient management, in particular to prevent the spread of resistant strains in the recent human H7N9 outbreak. We tailored a pyrosequencing assay to reveal neuraminidase R292K, a resistant marker found in one isolate from China, and demonstrated its performance in both sensitivity and specificity. In addition, a semi-nested polymerase chain reaction was applied, which enhanced the detection rate by at least 10-fold. We validated this assay by examining the marker in Taiwan's first imported human case and found R and K in quasispecies.
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan, ROC.
authors.forename: Chee-Keng
authors.initials: CK
authors.lastname: Mok
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC.
authors.forename: Shih-Cheng
authors.initials: SC
authors.lastname: Chang
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC; Department of Computer Science and Information Engineering, College of Engineering, Chang Gung University, Taoyuan, Taiwan, ROC.
authors.forename: Guang-Wu
authors.initials: GW
authors.lastname: Chen
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC.
authors.forename: Yu-Lun
authors.initials: YL
authors.lastname: Lo
authors.affiliation: Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan, ROC; Molecular Medicine Research Center, Chang Gung University, Taoyuan, Taiwan, ROC.
authors.forename: Shu-Jen
authors.initials: SJ
authors.lastname: Chen
authors.affiliation: Research and Diagnostics Center, Centers for Disease Control, Taipei, Taiwan, ROC.
authors.forename: Ho-Sheng
authors.initials: HS
authors.lastname: Wu
authors.affiliation: Research and Diagnostics Center, Centers for Disease Control, Taipei, Taiwan, ROC.
authors.forename: Ming-Tsan
authors.initials: MT
authors.lastname: Liu
authors.affiliation: Research and Diagnostics Center, Centers for Disease Control, Taipei, Taiwan, ROC.
authors.forename: Feng-Yee
authors.initials: FY
authors.lastname: Chang
authors.affiliation: Ministry of Health and Welfare, Executive Yuan, Taipei, Taiwan, ROC.
authors.forename: Tzou-Yien
authors.initials: TY
authors.lastname: Lin
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University and Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taiwan Centers for Disease Control Collaborating Laboratory of Virology, Taoyuan, Taiwan, ROC; Graduate Institute of Biomedical Sciences, College of Medicine, Chang Gung University, Taoyuan, Taiwan, ROC. Electronic address: srshih@mail.cgu.edu.tw.
authors.forename: Shin-Ru
authors.initials: SR
authors.lastname: Shih
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology*; D024882:Drug Resistance, Viral; D006801:Humans; D064766:Influenza A Virus, H7N9 Subtype / Q000187:drug effects* / Q000201:enzymology* / Q000302:isolation & purification; D007251:Influenza, Human / Q000821:virology; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D009439:Neuraminidase / Q000235:genetics*; D053139:Oseltamivir / Q000494:pharmacology*; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA; D013624:Taiwan; D014764:Viral Proteins / Q000235:genetics*
chemical_list: D000998:Antiviral Agents; D012367:RNA, Viral; D014764:Viral Proteins; D053139:Oseltamivir; C487630:NA protein, influenza A virus; D009439:Neuraminidase
keywords: Avian influenza A virus; H7N9; Oseltamivir-resistant marker; Pyrosequencing