title: Diagnostic performance of two highly multiplexed respiratory virus assays in a pediatric cohort.
abstract: BACKGROUND
  Rapid detection of respiratory viruses is important for management and infection control in hospitalized patients. Multiplex nucleic acid tests (NATs) have begun to replace conventional methods as gold standards for respiratory virus detection.
  
  
  OBJECTIVE
  To compare the performance of two large multiplex NATS, ResPlex II (RPII) and Respiratory Virus Surveillance kit with electrospray ionization mass spectrometry (RVS/MS) using nasopharyngeal aspirates (NPAs) from hospitalized children who had been tested previously with conventional methods.
  
  
  METHODS
  Stored residual NPAs (N=306) were tested concomitantly by RPII and RVS/MS. Alternate NATs were used to adjudicate discordant results.
  
  
  RESULTS
  More viruses were detected with multiplex NATs (RPII, 110; RVS/MS, 109) than conventional assays (86); diagnostic gain was primarily for fastidious viruses (coronaviruses and enteroviruses [EVs]/human rhinoviruses [HRVs]). Total positive and negative agreement between the multiplex NATs for all viruses detected was quite high (86% positive agreement, 99% negative agreement). Most individual viruses were detected with fairly equivalent accuracy by the multiplex NATs, except for adenoviruses (RPII sensitivity 40%) and human metapneumovirus (RVS/MS sensitivity 42%). RPII had the advantage of detecting EVs and HRVs, however, it demonstrated considerable EV/HRV cross-reactivity (29 HRV-positive specimens by real-time PCR were positive for EV by RPII and 21 specimens positive for HRV only by RT-PCR were dual positive for EV/HRV by RPII). RPII also had reduced sensitivity for HRV detection (in 36 specimens, HRV was detected by RT-PCR but not by RPII).
  
  
  CONCLUSIONS
  Both multiplex NATs were promising, but had notable limitations.
authors: Division of Medical Microbiology, The Johns Hopkins Medical Institutions, Baltimore, MD, United States.
authors: Michael S
authors: MS
authors: Forman
authors: Sonali
authors: S
authors: Advani
authors: Christina
authors: C
authors: Newman
authors: Charlotte A
authors: CA
authors: Gaydos
authors: Aaron M
authors: AM
authors: Milstone
authors: Alexandra
authors: A
authors: Valsamakis
mesh_terms: D002675:Child, Preschool; D015331:Cohort Studies; D006801:Humans; D007223:Infant; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009305:Nasopharynx / Q000821:virology; D012141:Respiratory Tract Infections / Q000175:diagnosis*; D012189:Retrospective Studies; D014773:Virology / Q000379:methods*; D014777:Virus Diseases / Q000175:diagnosis*