title: [Multiplex RT-PCR for the rapid detection of influenza virus types and subtypes].
abstract: OBJECTIVE
  To establish a sensitive and specific multiplex RT-PCR(MRT-PCR) for the simultaneous detection of influenza virus types and subtypes.
  
  
  METHODS
  Primers were designed from highly conserved region of the hemagglutinin(HA) gene of influenza H1N1H3N2 and B virus and MRT-PCR was performed. Additional two pairs of primers were designed to determine the N1 and N2 subtypes of neuramidinase NA of influenza H1N1 and H3N2 virus.
  
  
  RESULTS
  The fragments of HA gene of all types of influenza virus were amplified and there was no cross reaction. The sensitivity of detection of influenza H1N1 and H3N2 virus was 0.10 TCID50/50 microliter by the second PCR and that was 0.01 TCID50/50 microliter for influenza B virus. The NA gene of influenza H1N1 and H3N2 virus was also amplified by this method.
  
  
  CONCLUSIONS
  The sensitivity of detection of influenza virus from clinical patients' throat washing specimens by MRT-PCR was higher than that by MDCK cell culture or egg embryo isolation. This method was highly sensitive and timely for detection of influenza virus types and subtypes.
authors.affiliation: Zhejiang Center for Disease Control and Prevention, Hangzhou 310009, China.
authors.forename: Yi-yu
authors.initials: YY
authors.lastname: Lu
authors.forename: Ju-ying
authors.initials: JY
authors.lastname: Yan
authors.forename: Hai-yan
authors.initials: HY
authors.lastname: Mao
authors.forename: Yan
authors.initials: Y
authors.lastname: Sun
authors.forename: Min
authors.initials: M
authors.lastname: Zhou
authors.forename: Wen
authors.initials: W
authors.lastname: Shi
mesh_terms: D000818:Animals; D002460:Cell Line; D002642:Chick Embryo; D002645:Chickens; D017931:DNA Primers; D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics / Q000302:isolation & purification; D009980:Influenza A virus / Q000235:genetics* / Q000302:isolation & purification; D005585:Influenza in Birds / Q000821:virology; D007251:Influenza, Human / Q000821:virology; D012367:RNA, Viral / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*
chemical_list: D017931:DNA Primers; D012367:RNA, Viral