title: [The study of multiple RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses].
abstract: OBJECTIVE
  To establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses.
  
  
  METHODS
  Obtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance.
  
  
  RESULTS
  Successfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity.
  
  
  CONCLUSIONS
  Successfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.
authors.forename: Liang
authors.initials: L
authors.lastname: Yang
authors.forename: Xiao-mei
authors.initials: XM
authors.lastname: Zhang
authors.forename: Xiao-guang
authors.initials: XG
authors.lastname: Zhang
authors.forename: Jing
authors.initials: J
authors.lastname: Ma
authors.forename: Min
authors.initials: M
authors.lastname: Wang
authors.forename: Le-ying
authors.initials: LY
authors.lastname: Wen
authors.forename: Da-yan
authors.initials: DY
authors.lastname: Wang
authors.forename: Tian
authors.initials: T
authors.lastname: Bai
authors.forename: Yue-long
authors.initials: YL
authors.lastname: Shu
authors.forename: Yong-hua
authors.initials: YH
authors.lastname: Qian
authors.forename: Yi
authors.initials: Y
authors.lastname: Zeng
mesh_terms: D006801:Humans; D053118:Influenza A Virus, H1N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D053122:Influenza A Virus, H3N2 Subtype / Q000235:genetics / Q000302:isolation & purification*; D053124:Influenza A Virus, H5N1 Subtype / Q000235:genetics / Q000302:isolation & purification*; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity