title: Optimizing Virus Identification in Critically Ill Children Suspected of Having an Acute Severe Viral Infection.
abstract: OBJECTIVE
  Multiplex rapid viral tests and nasopharyngeal flocked swabs are increasingly used for viral testing in PICUs. This study aimed at evaluating how the sampling site and the type of diagnostic test influence test results in children with suspected severe viral infection.
  
  
  METHODS
  Prospective cohort study.
  
  
  METHODS
  PICUs at 21 tertiary pediatric referral centers in the United States.
  
  
  METHODS
  During the 2010-2011 and 2011-2012 influenza seasons, we enrolled children (6 mo to 17 yr old) who were suspected to have severe viral infection.
  
  
  METHODS
  We collected samples by using a standardized protocol for nasopharyngeal aspirate and nasopharyngeal flocked swabs in nonintubated patients and for endotracheal tube aspirate and nasopharyngeal flocked swabs in intubated patients.
  
  
  RESULTS
  Viral testing included a single reverse transcription-polymerase chain reaction influenza test and the GenMark Respiratory Viral Panel (20 viruses). We enrolled 90 endotracheally intubated and 133 nonintubated children. We identified influenza in 45 patients with reverse transcription-polymerase chain reaction testing; the multiplex panel was falsely negative for influenza in two patients (4.4%). Six patients (13.3%) had not been diagnosed with influenza in the PICU. Non-influenza viruses were identified in 172 of 223 children (77.1%). In nonintubated children, the same virus was identified by nasopharyngeal flocked swabs and nasopharyngeal aspirate in 133 of 183 paired samples (72.7%), with +nasopharyngeal aspirate/-nasopharyngeal flocked swabs in 32 of 183 paired samples (17.4%). In intubated children, the same virus was identified by nasopharyngeal flocked swabs and endotracheal tube aspirate in 67 of 94 paired samples (71.3%), with +nasopharyngeal flocked swabs/- endotracheal tube aspirate in 22 of 94 paired samples (23.4%). Most discrepancies were either adenovirus or rhinovirus in both groups.
  
  
  CONCLUSIONS
  Standardized specimen collection and sensitive diagnostic testing with a reverse transcription-polymerase chain reaction increased the identification of influenza in critically ill children. For most pathogenic viruses identified, results from nasopharyngeal flocked swabs agreed with those from nasopharyngeal or endotracheal aspirates.
authors.affiliation: 1Department of Anesthesia, Perioperative and Pain Medicine (Critical Care), Boston Children's Hospital, Boston, MA. 2Departments of Anaesthesia and Pediatrics, Harvard Medical School, Boston, MA. 3Department of Epidemiology, Boston University School of Public Health, Boston, MA. 4Tufts University School of Medicine, Boston, MA. 5Marshfield Clinic Research Foundation, Marshfield, WI. 6Department of Anesthesia and Critical Care, Children's Hospital of Philadelphia, Philadelphia, PA. 7Section of Critical Care Medicine, Department of Pediatrics, Texas Children's Hospital, Houston, TX. 8Pediatric Critical Care Division, Children's Hospital and Medical Center, Omaha, NE. 9Pediatric Critical Care Medicine, Maria Fareri Children's Hospital, Westchester, NY. 10Influenza Division, U.S. Centers for Disease Control and Prevention, Atlanta, GA. 11Battelle Memorial Institute, Atlanta, GA.
authors.forename: Adrienne G
authors.initials: AG
authors.lastname: Randolph
authors.forename: Anna A
authors.initials: AA
authors.lastname: Agan
authors.forename: Ryan F
authors.initials: RF
authors.lastname: Flanagan
authors.forename: Jennifer K
authors.initials: JK
authors.lastname: Meece
authors.forename: Julie C
authors.initials: JC
authors.lastname: Fitzgerald
authors.forename: Laura L
authors.initials: LL
authors.lastname: Loftis
authors.forename: Edward J
authors.initials: EJ
authors.lastname: Truemper
authors.forename: Simon
authors.initials: S
authors.lastname: Li
authors.forename: Jill M
authors.initials: JM
authors.lastname: Ferdinands
mesh_terms: D000208:Acute Disease; D000293:Adolescent; D002648:Child; D002675:Child, Preschool; D016638:Critical Illness; D005260:Female; D006801:Humans; D007223:Infant; D007251:Influenza, Human / Q000821:virology*; D008297:Male; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction; D009305:Nasopharynx / Q000821:virology; D009975:Orthomyxoviridae / Q000302:isolation & purification*; D011446:Prospective Studies; D060888:Real-Time Polymerase Chain Reaction; D012141:Respiratory Tract Infections / Q000821:virology*; D013048:Specimen Handling / Q000379:methods; D014777:Virus Diseases / Q000175:diagnosis* / Q000382:microbiology