title: Detection of low copies of drug-resistant influenza viral gene by a single-tube reaction using peptide nucleic acid as both PCR clamp and sensor probe.
abstract: Influenza virus infection causes endemics almost yearly and pandemics occasionally. Although antivirals are available for the clinical treatment of influenza virus infection, the emergence of a drug-resistant virus has reduced the effectiveness of therapy and prophylaxis. Therefore, the timely detection of drug-resistant influenza viruses is important. A single-tube reaction using peptide nucleic acid (PNA) as both a polymerase chain reaction (PCR) clamp and a sensor probe was established to detect the low numbers of copies of viral genes that carry the resistant marker. Influenza A H1N1 viruses resistant to a clinically used antiviral, amantadine, are selected for the experimental design. The PNA-mediated reverse transcription-PCR detected 10 copies/μL of RNA from the resistant strain among 2 × 10(4) copies/μL of RNA from the sensitive strain. A rapid and sensitive method was established for detecting low numbers of drug-resistant genes of the influenza virus. The assay would help to monitorthe emergence of adrug-resistant influenza virus.
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan, Taiwan; Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan, Taiwan; Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
authors.forename: Kuo-Chien
authors.initials: KC
authors.lastname: Tsao
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Chiuan-Chian
authors.initials: CC
authors.lastname: Chiou
authors.affiliation: Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Tai-Long
authors.initials: TL
authors.lastname: Chen
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan, Taiwan; Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan, Taiwan; Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taoyuan, Taiwan.
authors.forename: Chung-Guei
authors.initials: CG
authors.lastname: Huang
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan, Taiwan; Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan, Taiwan.
authors.forename: Erh-Fang
authors.initials: EF
authors.lastname: Hsieh
authors.affiliation: Research Center for Emerging Viral Infections, Chang Gung University, Taoyuan, Taiwan; Department of Medical Biotechnology and Laboratory Science, Chang Gung University, Taoyuan, Taiwan; Clinical Virology Laboratory, Chang Gung Memorial Hospital, Taoyuan, Taiwan. 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 / Q000235:genetics*; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000187:drug effects / Q000235:genetics; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology*; D020135:Peptide Nucleic Acids; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
chemical_list: D000998:Antiviral Agents; D020135:Peptide Nucleic Acids