title: Multiplex MassTag-PCR for respiratory pathogens in pediatric nasopharyngeal washes negative by conventional diagnostic testing shows a high prevalence of viruses belonging to a newly recognized rhinovirus clade.
abstract: BACKGROUND
  Respiratory infections are the most common infectious diseases in humans worldwide and are a leading cause of death in children less than 5 years of age.
  
  
  OBJECTIVE
  Identify candidate pathogens in pediatric patients with unexplained respiratory disease.
  
  
  METHODS
  Forty-four nasopharyngeal washes collected during the 2004-2005 winter season from pediatric patients with respiratory illnesses that tested negative for 7 common respiratory pathogens by culture and direct immunofluorescence assays were analyzed by MassTag-PCR. To distinguish human enteroviruses (HEV) and rhinoviruses (HRV), samples positive for picornaviruses were further characterized by sequence analysis.
  
  
  RESULTS
  Candidate pathogens were detected by MassTag PCR in 27 of the 44 (61%) specimens that previously were rated negative. Sixteen of these 27 specimens (59%) contained picornaviruses; of these 9 (57%) contained RNA of a recently discovered clade of rhinoviruses. Bocaviruses were detected in three patients by RT-PCR.
  
  
  CONCLUSIONS
  Our study confirms that multiplex MassTag-PCR enhances the detection of pathogens in clinical specimens, and shows that previously unrecognized rhinoviruses, that potentially form a species HRV-C, may cause a significant amount of pediatric respiratory disease.
authors.affiliation: Department of Pediatrics, The Children's Hospital, University of Colorado Denver School of Medicine, Aurora, CO, USA.
authors.forename: Samuel R
authors.initials: SR
authors.lastname: Dominguez
authors.forename: Thomas
authors.initials: T
authors.lastname: Briese
authors.forename: Gustavo
authors.initials: G
authors.lastname: Palacios
authors.forename: Jeffrey
authors.initials: J
authors.lastname: Hui
authors.forename: Joseph
authors.initials: J
authors.lastname: Villari
authors.forename: Vishal
authors.initials: V
authors.lastname: Kapoor
authors.forename: Rafal
authors.initials: R
authors.lastname: Tokarz
authors.forename: Mary P
authors.initials: MP
authors.lastname: Glodé
authors.forename: Marsha S
authors.initials: MS
authors.lastname: Anderson
authors.forename: Christine C
authors.initials: CC
authors.lastname: Robinson
authors.forename: Kathryn V
authors.initials: KV
authors.lastname: Holmes
authors.forename: W Ian
authors.initials: WI
authors.lastname: Lipkin
mesh_terms: D000208:Acute Disease; D002675:Child, Preschool; D004770:Enterovirus / Q000302:isolation & purification; D004769:Enterovirus Infections / Q000453:epidemiology / Q000821:virology; D005260:Female; D006801:Humans; D007223:Infant; D008297:Male; D009305:Nasopharynx / Q000821:virology*; D010850:Picornaviridae Infections / Q000453:epidemiology / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods; D015995:Prevalence; D012141:Respiratory Tract Infections / Q000453:epidemiology / Q000821:virology; D012229:Rhinovirus / Q000145:classification / Q000302:isolation & purification; D014777:Virus Diseases / Q000453:epidemiology / Q000821:virology