title: Multiplex SYBR Green Real-Time PCR Assay for Detection of Respiratory Viruses.
abstract: BACKGROUND
  It is often difficult for a physician to distinguish between viral and bacterial causes of respiratory infections and this may result in overuse of antibiotics. In many cases of community-acquired respiratory infections, clinicians treat patients empirically. The development of molecular methods for direct detection of viruses has been progressed recently.
  
  
  OBJECTIVE
  The objective of this study was recognizing the panel of respiratory RNA viruses by multiplex SYBR Green real-time polymerase chain reaction (PCR).
  
  
  METHODS
  Randomized 172 influenza-negative respiratory specimens of all age groups of hospitalized patients were collected. After RNA extraction, cDNA was synthesized. Three SYBR Green multiplex real-time PCR assays were developed for simultaneous detection of 12 respiratory RNA viruses. Each set of multiplex methods detected four viruses, the first set: respiratory syncytial virus, human metapneumovirus, rhinovirus, enterovirus; the second set: parainfluenza viruses 1 - 4 (PIV1-4); the third set: coronaviruses NL63, 229E, severe acute respiratory syndrome (SARS), and OC43.
  
  
  RESULTS
  Application of the multiplex SYBR Green real-time PCR in clinical samples from 172 patients in a one-year study resulted in detection of 19 (11.04%) PIV3, 9 (5.23%) PIV4, and 1 (0.58%) coronavirus NL63. All the positive samples were detected during December to March (2011 - 2012).
  
  
  CONCLUSIONS
  Multiplex SYBR Green real-time PCR is a rapid and relatively inexpensive method for detection of respiratory viruses.
authors.affiliation: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Mozhdeh
authors.initials: M
authors.lastname: Sultani
authors.affiliation: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Talat
authors.initials: T
authors.lastname: Mokhtari Azad
authors.affiliation: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Mohammadreza
authors.initials: M
authors.lastname: Eshragian
authors.affiliation: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Azadeh
authors.initials: A
authors.lastname: Shadab
authors.affiliation: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Naseri
authors.affiliation: Department of Infectious Disease, Yasuj University of Medical Sciences, Yasuj, IR Iran.
authors.forename: Owrang
authors.initials: O
authors.lastname: Eilami
authors.affiliation: Virology Department, School of Public Health, Tehran University of Medical Sciences, Tehran, IR Iran.
authors.forename: Jila
authors.initials: J
authors.lastname: Yavarian
keywords: Real-Time PCR; Respiratory Syncytial Viruses; SYBR Green