title: Influenza A/B virus detection and influenza A virus subtyping with emphasis on the novel H7N9 virus by using multiplex real-time RT-PCR.
abstract: Infections of the novel avian influenza A H7N9 virus cause severe respiratory diseases and death. In this study, to develop highly sensitive methods for differentially detecting the H7N9 virus, multiplex and singular real-time reverse transcription polymerase chain reaction (RT-PCR) assays were established and examined by targeting the H7 and N9 genes of the H7N9 virus. Furthermore, an additional multiplex assay combining previous real time RT-PCR designs was established to subtype the pandemic H1N1, H3, and H5 influenza viruses. Applying the proposed assay system to analyze 100 clinical specimens collected from respiratory infection cases identified influenza A viruses (pandemic H1N1 and H3) in 23 samples. It has been demonstrated that other common respiratory viruses will not be detected by using this platform.
authors: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Department of Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taiwan.
authors: Rei-Lin
authors: RL
authors: Kuo
authors: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.
authors: Shu-Li
authors: SL
authors: Yang
authors: Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.
authors: Yi-Chun
authors: YC
authors: Liu
authors: Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.
authors: Lu-Ting
authors: LT
authors: Chen
authors: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan.
authors: Chee-Keng
authors: CK
authors: Mok
authors: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan.
authors: Shu-Ming
authors: SM
authors: Kuo
authors: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Department of Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taiwan; Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan.
authors: Shin-Ru
authors: SR
authors: Shih
authors: Research Center for Emerging Viral Infections, College of Medicine, Chang Gung University, Taiwan; Department of Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taiwan; Department of Laboratory Medicine, Linkou Chang Gung Memorial Hospital, Taoyuan, Taiwan. Electronic address: kctsao@cgmh.org.tw.
authors: Kuo-Chien
authors: KC
authors: Tsao
mesh_terms: D006801:Humans; D009980:Influenza A virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D009981:Influenza B virus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
keywords: Influenza A H7N9 virus; Multiplex real-time reverse transcription polymerase chain reaction (RT-PCR); Rapid and sensitive detection