A multiplex RT-PCR for the detection of parainfluenza viruses 1-3 in clinical samples

J Virol Methods. 1999 Sep;82(1):9-18. doi: 10.1016/s0166-0934(99)00073-7.


Parainfluenza viruses (PIV) are an important cause of respiratory morbidity. Conventional diagnostic methods for detection of PIV are time consuming or lack sensitivity. A multiplex PCR that detects PIV 1-3 was developed using novel primers for PIV viruses 1 and 2 and primers for PIV 3 described previously. Following RNA extraction a single multiplex reverse transcription was undertaken using antisense primers specific for each virus type. This was followed by a 40-cycle multiplex PCR using primers directed towards the haemagglutinin-neuraminidase coding region of each virus type. Products were probed with type-specific fluorescein labelled internal probes and detected by chemiluminescence. Cultured PIV viruses were detectable to a sensitivity of 1 TCID50. The technique was applied to 57 nasal aspirates taken from children presenting with various acute respiratory conditions and analysed previously by culture, immunofluorescence and/or serology. It was possible to detect PIV 1, 2 or 3 in 13/13 samples found previously positive for PIV by tissue culture, 13/15 found previously positive by immunofluorescence and 6/10 that coincided with positive serology. None of the samples found previously positive for other viruses (26) or negative to virus detection (6) were found positive by RT-PCR. It is concluded that this method is as sensitive as combined immunofluorescence and tissue culture for the detection of the PIV viruses 1-3 and should be useful for rapid diagnosis of PIV 1-3 infections.

MeSH terms

  • Animals
  • Cell Line
  • Child
  • Croup / pathology
  • Croup / virology*
  • Culture Techniques / methods
  • Fluorescent Antibody Technique
  • Humans
  • Macaca mulatta
  • Parainfluenza Virus 1, Human / genetics
  • Parainfluenza Virus 1, Human / isolation & purification*
  • Parainfluenza Virus 2, Human / genetics
  • Parainfluenza Virus 2, Human / isolation & purification*
  • Parainfluenza Virus 3, Human / genetics
  • Parainfluenza Virus 3, Human / isolation & purification*
  • RNA, Viral / analysis
  • Respirovirus Infections / pathology
  • Respirovirus Infections / virology*
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity


  • RNA, Viral