title: Role of real-time reverse transcription polymerase chain reaction for detection of respiratory viruses in critically ill children with respiratory disease: Is it time for a change in algorithm?
abstract: OBJECTIVE
  To identify the respiratory viral pathogens associated with acute lower respiratory tract infection in critically ill pediatric patients by using real-time reverse transcription-polymerase chain reaction, and compare results with those of direct fluorescence antibody assay testing.
  
  
  METHODS
  Observational cohort study.
  
  
  METHODS
  Pediatric intensive care unit at a tertiary care academic hospital.
  
  
  METHODS
  Pediatric patients admitted to the pediatric intensive care unit with severe respiratory symptoms consistent with viral lower respiratory tract infection.
  
  
  METHODS
  None.
  
  
  METHODS
  Respiratory samples of pediatric patients admitted to the pediatric intensive care unit with severe respiratory symptoms between January 2008 and July 2009 were tested with direct fluorescence antibody assay and real-time reverse transcription-polymerase chain reaction.
  
  
  RESULTS
  At least one viral agent was detected in 70.5% of specimens by real-time reverse transcription-polymerase chain reaction and in 16.5% by direct fluorescence antibody assay (p < .001). Real-time reverse transcription-polymerase chain reaction increased the total viral yield five-fold compared to direct fluorescence antibody assay. Rhinovirus was the most commonly identified virus (41.6%). For viruses included in the direct fluorescence antibody assay panel, direct fluorescence antibody assay had a sensitivity of 0.42 (95% confidence interval 0.25-0.61) and a specificity of 1 (95% confidence interval 0.86-1.00) compared with real-time reverse transcription-polymerase chain reaction. Coinfections were not uncommon, in particular with rhinovirus, and these patients tended to have higher mortality.
  
  
  CONCLUSIONS
  Direct fluorescence antibody assay testing is a suboptimal method for the detection of respiratory viruses in critically ill children with lower respiratory tract infection. Given the importance of a prompt and accurate viral diagnosis for this group of patients, we suggest that real-time reverse transcription-polymerase chain reaction becomes part of the routine diagnostic algorithm in critically ill children when a viral etiology is suspected, even if conventional tests yield a negative result.
authors: Department of Pediatrics, Division of Pediatric Critical Care Medicine, University of California at San Francisco, San Francisco, CA, USA.
authors: Angela
authors: A
authors: Aramburo
authors: Sandrijn
authors: S
authors: van Schaik
authors: Janice
authors: J
authors: Louie
authors: Erica
authors: E
authors: Boston
authors: Sharon
authors: S
authors: Messenger
authors: Carolyn
authors: C
authors: Wright
authors: W
authors: W
authors: Lawrence Drew
mesh_terms: D000208:Acute Disease; D000465:Algorithms; D002675:Child, Preschool; D015331:Cohort Studies; D016638:Critical Illness; D005260:Female; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D007223:Infant; D007251:Influenza, Human / Q000175:diagnosis / Q000821:virology; D008297:Male; D009975:Orthomyxoviridae / Q000302:isolation & purification; D010850:Picornaviridae Infections / Q000175:diagnosis / Q000821:virology; D012328:RNA Viruses / Q000302:isolation & purification*; D060888:Real-Time Polymerase Chain Reaction; D018357:Respiratory Syncytial Virus Infections / Q000175:diagnosis / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000302:isolation & purification; D012141:Respiratory Tract Infections / Q000175:diagnosis / Q000821:virology*; D018108:Respirovirus / Q000302:isolation & purification; D010253:Respirovirus Infections / Q000175:diagnosis / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012229:Rhinovirus / Q000302:isolation & purification; D012680:Sensitivity and Specificity