Evaluation of a PCR/ESI-MS platform to identify respiratory viruses from nasopharyngeal aspirates

J Med Virol. 2015 Nov;87(11):1867-71. doi: 10.1002/jmv.24262. Epub 2015 May 20.

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.

Keywords: diagnosis; direct immunofluorescent assay (DFA); electrospray ionization mass spectrometry (ESI/MS); respiratory tract infections; virus.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Child, Preschool
  • Clinical Laboratory Techniques / methods
  • Female
  • Fluorescent Antibody Technique, Direct
  • Humans
  • Infant
  • Male
  • Nasopharynx / virology*
  • Polymerase Chain Reaction / methods*
  • Respiratory Tract Infections / diagnosis*
  • Respiratory Tract Infections / virology
  • Sensitivity and Specificity
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Virology / methods
  • Virus Diseases / diagnosis*
  • Virus Diseases / virology
  • Viruses / classification*
  • Viruses / isolation & purification*