Evaluation of a polymerase chain reaction-electrospray ionization time-of-flight mass spectrometry for the detection and subtyping of influenza viruses in respiratory specimens

聚合酶链式反应-电喷雾电离飞行时间质谱法在呼吸道标本中流感病毒检测和分型中的应用评价

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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. OBJECTIVES: To evaluate the performance of the PLEX-ID flu detection™ kit for detecting Influenza viruses by comparison with the multiplex RespiFinder(®) Kit (PathoFinder). STUDY DESIGN: 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.

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