title: A novel high-speed droplet-polymerase chain reaction can detect human influenza virus in less than 30 min.
abstract: BACKGROUND
  The polymerase chain reaction (PCR) has been widely used for diagnosis of infection. Rapid detection of influenza virus is useful for therapeutic decisions. We attempted to develop a novel assay by real-time droplet-PCR machine for influenza virus.
  
  
  METHODS
  RNA extracted from nasal swabs or primary swabs pretreated only were used for PCR analyses. We evaluated reaction time, amplification efficiency, sensitivity, and specificity of the novel droplet-PCR.
  
  
  RESULTS
  The reaction time of the novel droplet-PCR was 28 min, whereas that of PCR using the conventional PCR machine was 80 min. The standard curve constructed from the amplification plots by the novel droplet-PCR was: y=-3.6x+42.9; that by PCR using the conventional PCR machine was: y=-3.5x+37.8. The sensitivity and specificity of the novel droplet-PCR were 86.7% and 91.7% for the influenza A and 100.0% and 100.0% for the influenza B, respectively. The novel droplet-PCR provided the specific amplification when using primary swabs without RNA extraction.
  
  
  CONCLUSIONS
  Our novel droplet-PCR markedly reduced the reaction time while showing same reactivity as that by PCR using the conventional PCR machine. Thus, the novel droplet-PCR assay can be used as a rapid assay for detection of influenza virus.
authors: Department of Laboratory Medicine, Shinshu University Hospital, 3-1-1 Asahi, Matsumoto 390-8621, Japan.
authors: Kazuyuki
authors: K
authors: Matsuda
authors: Akemi
authors: A
authors: Yamaguchi
authors: Chiaki
authors: C
authors: Taira
authors: Akane
authors: A
authors: Sueki
authors: Hiroshi
authors: H
authors: Koeda
authors: Fumio
authors: F
authors: Takagi
authors: Mitsutoshi
authors: M
authors: Sugano
authors: Takayuki
authors: T
authors: Honda
mesh_terms: D006801:Humans; D009980:Influenza A virus / Q000235:genetics*; D009981:Influenza B virus / Q000235:genetics*; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology*; D009296:Nasal Cavity / Q000821:virology; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D012367:RNA, Viral