title: [One-step multiplex RT-PCR for rapid screening of type A, B and novel A (H1N1) influenza viruses].
abstract: OBJECTIVE
  To developed a multiplex RT-PCR assay for simultaneous screening of type A, B and novel A (H1N1) influenza viruses.
  
  
  METHODS
  Two pairs of universal primers in were designed for amplifying the M gene and NS gene of type A and B influenza viruses, respectively. A pair of specific primers of HA gene was designed to detect novel A (H1N1) influenza virus. A one-step method was used to establish the multiplex RT-PCR system. A blinded experiment was carried out to validate the accuracy of this assay in comparison with the results of real-time fluorescence RT-PCR. The clinical practicability and efficacy of this assay was also evaluated.
  
  
  RESULTS
  The RT-PCR products were analyzed using agarose gel electrophoresis, which yielded distinct bands of the target fragments without non-specific reactions, suggesting the high efficiency and specificity of the multiplex RT-PCR. Blinded study of 50 samples demonstrated a concordance rate of 100%.
  
  
  CONCLUSIONS
  This multiplex RT-PCR assay allows one-step simultaneous detection of type A, B and novel A (H1N1) influenza viruses rapidly and accurately, and provides a valuable low-cost screening technique for influenza epidemic monitoring and early diagnosis.
authors.affiliation: Zhuhai Entry-Exit Inspection and Quarantine Bureau, Zhuhai 519020, China.
authors.forename: Qiu-hua
authors.initials: QH
authors.lastname: Mo
authors.forename: Cui-lan
authors.initials: CL
authors.lastname: Yang
authors.forename: Ji-can
authors.initials: JC
authors.lastname: Lin
authors.forename: Hua
authors.initials: H
authors.lastname: Tan
authors.forename: Cheng-ning
authors.initials: CN
authors.lastname: Tu
authors.forename: Li-qing
authors.initials: LQ
authors.lastname: Ye
authors.forename: Zhi-ming
authors.initials: ZM
authors.lastname: Liu
authors.forename: Jian
authors.initials: J
authors.lastname: DU
authors.forename: Hong
authors.initials: H
authors.lastname: Sun
authors.forename: Ze
authors.initials: Z
authors.lastname: Yang
mesh_terms: D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics* / Q000302:isolation & purification*; D009981:Influenza B virus / Q000235:genetics* / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013997:Time Factors; D014763:Viral Matrix Proteins / Q000235:genetics; D017361:Viral Nonstructural Proteins / Q000235:genetics
chemical_list: D014763:Viral Matrix Proteins; D017361:Viral Nonstructural Proteins