title: Simplified screening method for detecting oseltamivir resistant pandemic influenza A (H1N1) 2009 virus by a RT-PCR/restriction fragment length polymorphism assay.
abstract: The sudden emergence of the pandemic influenza A (H1N1) 2009 virus in early 2009 has resulted in a rapid transmission of this virus worldwide. Within a short time span, sporadic cases infected with this virus that shows oseltamivir resistance have also been reported. These resistant viruses have an amino acid change from histidine to tyrosine at position 275 (H275Y) of the neuraminidase gene. In this study, a reverse transcriptase PCR/restriction fragment length polymorphism (RT-PCR/RFLP) assay was developed to detect the H275Y mutation. Resistant and sensitive viruses could be differentiated using the RFLP patterns. This RT-PCR/RFLP assay is a simple method and also very specific and sensitive for detecting the H275Y mutation of pandemic influenza A (H1N1) 2009 viruses, and can be used in resource-limited settings.
authors.affiliation: Department of Virology, Tohoku University Graduate School of Medicine, Sendai, Miyagi, Japan.
authors.forename: Nao
authors.initials: N
authors.lastname: Nukiwa
authors.forename: Akira
authors.initials: A
authors.lastname: Suzuki
authors.forename: Yuki
authors.initials: Y
authors.lastname: Furuse
authors.forename: Kozue
authors.initials: K
authors.lastname: Shimabukuro
authors.forename: Takashi
authors.initials: T
authors.lastname: Odagiri
authors.forename: Irona
authors.initials: I
authors.lastname: Khandaker
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Oshitani
mesh_terms: D019943:Amino Acid Substitution; D000998:Antiviral Agents / Q000494:pharmacology* / Q000627:therapeutic use; D024882:Drug Resistance, Viral / Q000235:genetics*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects* / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000188:drug therapy / Q000821:virology; D008826:Microbial Sensitivity Tests; D009439:Neuraminidase / Q000235:genetics; D053139:Oseltamivir / Q000494:pharmacology* / Q000627:therapeutic use; D012150:Polymorphism, Restriction Fragment Length; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA
chemical_list: D000998:Antiviral Agents; D053139:Oseltamivir; D009439:Neuraminidase