title: [Simultaneous detection of human parainfluenza viruses 1, 2, 3 by multiplex real-time reverse transcription-polymerase chain reaction with LNA probes].
abstract: OBJECTIVE
  Human parainfluenza virus (HPIV) types 1, 2 and 3 are major viral pathogens responsible for upper and lower respiratory tract infections. In this study, a real-time RT-PCR was developed using multiplex primers-probe (HPIV-1, 2, 3) for the simultaneous detection of both HPIV1, HPIV2 and HPIV3 genomes.
  
  
  METHODS
  Optimal primers and probes were designed using specialized software. The conditions for multiplex real-time RT-PCR had been optimized. The synthesis of RNA standards of HPIV1, 2, 3 were used a T7 RNA polymerase. Check the specificity sensitivities and stability of one step RT-PCR assay.
  
  
  RESULTS
  Obtained in a 10-fold dilution series assay demonstrate a high sensitivity of the assay with a lowest detection limit of 10 copies for HPIV1, 100 copies for HPIV2 and 100 copies for HPIV3.
  
  
  CONCLUSIONS
  The assays demonstrates an improved sensitivity and scope of detecting HPIV1, 2, 3 viruses relative to routine antigen detection assays while the quantitative utility may facilitate investigation of the pre-diagnosis and respiratory virus pathogenesis.
authors.affiliation: Institute for Viral Disease Control and Prevention, Chinese CDC, Beijing 102206, China.
authors.forename: Yi-Xin
authors.initials: YX
authors.lastname: Ji
authors.forename: Nai-Ying
authors.initials: NY
authors.lastname: Mao
authors.forename: Huan-Huan
authors.initials: HH
authors.lastname: Wang
authors.forename: Zheng-De
authors.initials: ZD
authors.lastname: Xie
authors.forename: Wen-Bo
authors.initials: WB
authors.lastname: Xu
mesh_terms: D006801:Humans; D009841:Oligonucleotides / Q000235:genetics*; D010222:Parainfluenza Virus 1, Human / Q000302:isolation & purification*; D010223:Parainfluenza Virus 2, Human / Q000302:isolation & purification*; D010224:Parainfluenza Virus 3, Human / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
chemical_list: D009841:Oligonucleotides; C477371:locked nucleic acid