title: Evaluation of a multiplex real-time PCR assay for the detection of respiratory viruses in clinical specimens.
abstract: BACKGROUND
  In this study, we evaluated the analytical performance and clinical potential of a one-step multiplex real-time PCR assay for the simultaneous detection of 14 types of respiratory viruses using the AdvanSure RV real-time PCR Kit (LG Life Sciences, Korea).
  
  
  METHODS
  Three hundred and twenty clinical specimens were tested with the AdvanSure RV real-time PCR Kit and conventional multiplex reverse transcription (RT)-PCR assay. The assay results were analyzed and the one-step AdvanSure RV real-time PCR Kit was compared with the conventional multiplex RT-PCR assay with respect to the sensitivity and specificity of the detection of respiratory viruses.
  
  
  RESULTS
  The limit of detection (LOD) was 1.31 plaque-forming units (PFU)/mL for human rhinoviruses (hRVs), 4.93 PFU/mL for human coronavirus HCoV-229E/NL63, 2.67 PFU/mL for human coronavirus HCoV-OC43, 18.20 PFU/mL for parainfluenza virus 1 (PIV)-1, 24.57 PFU/mL for PIV-2, 1.73 PFU/mL for PIV-3, 1.79 PFU/mL for influenza virus group (Flu) A, 59.51 PFU/mL for FluB, 5.46 PFU/mL for human respiratory syncytial virus (hRSV)-A, 17.23 PFU/mL for hRSV-B, 9.99 PFU/mL for human adenovirus (ADVs). The cross-reactivity test for this assay against 23 types of non-respiratory viruses showed negative results for all viruses tested. The agreement between the one-step AdvanSure multiplex real-time PCR assay and the conventional multiplex RT-PCR assay was 98%.
  
  
  CONCLUSIONS
  The one-step AdvanSure RV multiplex real-time PCR assay is a simple assay with high potential for specific, rapid and sensitive laboratory diagnosis of respiratory viruses compared to conventional multiplex RT-PCR.
authors.affiliation: Department of Laboratory Medicine, Dankook University College of Medicine, Cheonan, Korea.
authors.forename: Insoo
authors.initials: I
authors.lastname: Rheem
authors.forename: Joowon
authors.initials: J
authors.lastname: Park
authors.forename: Tae-Hyun
authors.initials: TH
authors.lastname: Kim
authors.forename: Jong Wan
authors.initials: JW
authors.lastname: Kim
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D017934:Coronavirus / Q000235:genetics; D004279:DNA, Viral / Q000032:analysis*; D006801:Humans; D008171:Lung Diseases / Q000175:diagnosis / Q000821:virology*; D060885:Multiplex Polymerase Chain Reaction; D009975:Orthomyxoviridae / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis*; D011933:Reagent Kits, Diagnostic; D018113:Respiratory Syncytial Virus, Human / Q000235:genetics; D018108:Respirovirus / Q000235:genetics; D012229:Rhinovirus / Q000235:genetics
chemical_list: D004279:DNA, Viral; D012367:RNA, Viral; D011933:Reagent Kits, Diagnostic
keywords: Limit of detection; Multiplex real-time PCR; Respiratory virus