title: Evaluation of a polymerase chain reaction-electrospray ionization time-of-flight mass spectrometry for the detection and subtyping of influenza viruses in respiratory specimens.
abstract: BACKGROUND
  PCR coupled to electrospray ionization mass spectrometry technology (PCR/ESI-TOF-MS) (PLEX-ID system, Abbott Ibis Biosciences) was developed to characterize microbial pathogens.
  
  
  OBJECTIVE
  To evaluate the performance of the PLEX-ID flu detection™ kit for detecting Influenza viruses by comparison with the multiplex RespiFinder(®) Kit (PathoFinder).
  
  
  METHODS
  Acute-phase respiratory samples (n=293) were analysed for this purpose. A subpopulation of influenza type A positive samples, identified with the RespiFinder(®) kit (n=64), were subtyped with the RealTime ready Inf A/H1N1 Detection Set(®) (Roche Molecular Diagnostics) and results were compared to the PLEX-ID Flu Detection™ kit.
  
  
  RESULTS
  274 samples gave concordant results (93.5%, p<0.0001): 65 influenza A-positive, 18 influenza B-positive and 191 negative samples. Of these, 7 samples were PLEX-ID positive/RespiFinder(®) negative (5 influenza A and 2 influenza B) and 12 were PLEX-ID positive/RespiFinder(®) negative (10 influenza A and 2 influenza B). PLEX-ID showed one sample as an influenza A and B co-infection while the RespiFinder(®) assay showed it to be influenza A-positive. The sensitivity, specificity, positive and negative predictive values of the PLEX-ID™ system were 87.4%, 96.5%, 92.2% and 94.1% respectively. Thirteen of 19 discordant samples available for retesting were investigated further with the Anyplex™II RV16 Detection kit (Seegene): seven were RespiFinder(®) concordant, while six were PLEX-ID™ concordant. Subtyping of 61/64 influenza A samples was concordant (95.3%): 55 were H1N1pdm09 and six were non-H1N1pdm09. Three samples gave negative PLEX-ID™ results (one H1N1pdm09 and two non-H1N1pdm09).
  
  
  CONCLUSIONS
  PCR/ESI-TOF-MS technology showed good diagnostic performances to detect and subtype influenza viruses.
authors.affiliation: Department of Virology, CH Toulouse, 330 Avenue de Grande Bretagne, TSA 40031, 31059 Toulouse Cedex 09, France. mengelle.c@chu-toulouse.fr
authors.forename: Catherine
authors.initials: C
authors.lastname: Mengelle
authors.forename: Jean-Michel
authors.initials: JM
authors.lastname: Mansuy
authors.forename: Isabelle
authors.initials: I
authors.lastname: Da Silva
authors.forename: Jean-Luc
authors.initials: JL
authors.lastname: Guerin
authors.forename: Jacques
authors.initials: J
authors.lastname: Izopet
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D012634:Bodily Secretions / Q000821:virology; D002648:Child; D002675:Child, Preschool; D060085:Coinfection / Q000175:diagnosis / Q000821:virology; D005260:Female; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D007251:Influenza, Human / Q000175:diagnosis* / Q000821:virology*; D008297:Male; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009975:Orthomyxoviridae / Q000737:chemistry / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012137:Respiratory System / Q000821:virology; D012680:Sensitivity and Specificity; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014773:Virology / Q000379:methods; D055815:Young Adult
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