title: Evaluation of a PCR/ESI-MS platform to identify respiratory viruses from nasopharyngeal aspirates.
abstract: Acute respiratory tract infection is a major cause of morbidity and mortality worldwide, particularly in infants and young children. High-throughput, accurate, broad-range tools for etiologic diagnosis are critical for effective epidemic control. In this study, the diagnostic capacities of an Ibis platform based on the PCR/ESI-MS assay were evaluated using clinical samples. Nasopharyngeal aspirates (NPAs) were collected from 120 children (<5 years old) who were hospitalized with lower respiratory tract infections between November 2010 and October 2011. The respiratory virus detection assay was performed using the PCR/ESI-MS assay and the DFA. The discordant PCR/ESI-MS and DFA results were resolved with RT-PCR plus sequencing. The overall agreement for PCR/ESI-MS and DFA was 98.3% (118/120). Compared with the results from DFA, the sensitivity and specificity of the PCR/ESI-MS assay were 100% and 97.5%, respectively. The PCR/ESI-MS assay also detected more multiple virus infections and revealed more detailed subtype information than DFA. Among the 12 original specimens with discordant results between PCR/ESI-MS and DFA, 11 had confirmed PCR/ESI-MS results. Thus, the PCR/ESI-MS assay is a high-throughput, sensitive, specific and promising method to detect and subtype conventional viruses in respiratory tract infections and allows rapid identification of mixed pathogens.
authors.affiliation: Department of Laboratory Medicine, Huashan Hospital of Fudan University, Shanghai, China.|Department of Center Laboratory, Jingan District Center Hospital of Shanghai, Shanghai, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Lin
authors.affiliation: Department of Medical Microbiology and Parasitology, Institutes of Biomedical Sciences, Shanghai Medical College of Fudan University, Shanghai, China.
authors.forename: Yongfeng
authors.initials: Y
authors.lastname: Fu
authors.affiliation: Department of Clinical Laboratory Center, Children's Hospital of Fudan University, Shanghai, China.
authors.forename: Menghua
authors.initials: M
authors.lastname: Xu
authors.affiliation: Department of Clinical Laboratory Center, Children's Hospital of Fudan University, Shanghai, China.
authors.forename: Liyun
authors.initials: L
authors.lastname: Su
authors.affiliation: Department of Clinical Laboratory Center, Children's Hospital of Fudan University, Shanghai, China.
authors.forename: Lingfeng
authors.initials: L
authors.lastname: Cao
authors.affiliation: Department of Clinical Laboratory Center, Children's Hospital of Fudan University, Shanghai, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Xu
authors.affiliation: Department of Medical Microbiology and Parasitology, Institutes of Biomedical Sciences, Shanghai Medical College of Fudan University, Shanghai, China.
authors.forename: Xunjia
authors.initials: X
authors.lastname: Cheng
mesh_terms: D002675:Child, Preschool; D019411:Clinical Laboratory Techniques / Q000379:methods; D005260:Female; D019085:Fluorescent Antibody Technique, Direct; D006801:Humans; D007223:Infant; D008297:Male; D009305:Nasopharynx / Q000821:virology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D014773:Virology / Q000379:methods; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology; D014780:Viruses / Q000145:classification* / Q000302:isolation & purification*
keywords: diagnosis; direct immunofluorescent assay (DFA); electrospray ionization mass spectrometry (ESI/MS); respiratory tract infections; virus