title: Rapid and generic identification of influenza A and other respiratory viruses with mass spectrometry.
abstract: The rapid identification of existing and emerging respiratory viruses is crucial in combating outbreaks and epidemics. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) is a rapid and reliable identification method in bacterial diagnostics, but has not been used in virological diagnostics. Mass spectrometry systems have been investigated for the identification of respiratory viruses. However, sample preparation methods were laborious and time-consuming. In this study, a reliable and rapid sample preparation method was developed allowing identification of cultured respiratory viruses. Tenfold serial dilutions of ten cultures influenza A strains, mixed samples of influenza A virus with human metapneumovirus or respiratory syncytial virus, and reconstituted clinical samples were treated with the developed sample preparation method. Subsequently, peptides were subjected to MALDI-TOF MS and liquid chromatography tandem mass spectrometry (LC-MS/MS). The influenza A strains were identified to the subtype level within 3h with MALDI-TOF MS and 6h with LC-MS/MS, excluding the culturing time. The sensitivity of LC-MS/MS was higher compared to MALDI-TOF MS. In addition, LC-MS/MS was able to discriminate between two viruses in mixed samples and was able to identify virus from reconstituted clinical samples. The development of an improved and rapid sample preparation method allowed generic and rapid identification of cultured respiratory viruses by mass spectrometry.
authors.affiliation: Netherlands Organization for Applied Scientific Research TNO, Department of CBRN Protection, Lange Kleiweg 137, 2288 GJ Rijswijk, The Netherlands.
authors.forename: Joanna A
authors.initials: JA
authors.lastname: Majchrzykiewicz-Koehorst
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands.
authors.forename: Esther
authors.initials: E
authors.lastname: Heikens
authors.affiliation: Netherlands Organization for Applied Scientific Research TNO, Department of CBRN Protection, Lange Kleiweg 137, 2288 GJ Rijswijk, The Netherlands.
authors.forename: Hein
authors.initials: H
authors.lastname: Trip
authors.affiliation: Netherlands Organization for Applied Scientific Research TNO, Department of CBRN Protection, Lange Kleiweg 137, 2288 GJ Rijswijk, The Netherlands.
authors.forename: Albert G
authors.initials: AG
authors.lastname: Hulst
authors.affiliation: Netherlands Organization for Applied Scientific Research TNO, Department of CBRN Protection, Lange Kleiweg 137, 2288 GJ Rijswijk, The Netherlands.
authors.forename: Ad L
authors.initials: AL
authors.lastname: de Jong
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands.
authors.forename: Marco C
authors.initials: MC
authors.lastname: Viveen
authors.affiliation: Netherlands Organization for Applied Scientific Research TNO, Department of CBRN Protection, Lange Kleiweg 137, 2288 GJ Rijswijk, The Netherlands.
authors.forename: Norbert J A
authors.initials: NJ
authors.lastname: Sedee
authors.affiliation: Expert Centre Force Health Protection, Health Care Division, Support Command, Ministry of Defence, Korte Molenweg 3, Building 37, 3941 PW Doorn, The Netherlands.
authors.forename: Jan
authors.initials: J
authors.lastname: van der Plas
authors.affiliation: Department of Medical Microbiology, University Medical Center Utrecht, Heidelberglaan 100, 3584 CX Utrecht, The Netherlands.
authors.forename: Frank E J
authors.initials: FE
authors.lastname: Coenjaerts
authors.affiliation: Netherlands Organization for Applied Scientific Research TNO, Department of CBRN Protection, Lange Kleiweg 137, 2288 GJ Rijswijk, The Netherlands. Electronic address: armand.paauw@tno.nl.
authors.forename: Armand
authors.initials: A
authors.lastname: Paauw
mesh_terms: D006801:Humans; D009980:Influenza A virus; D007251:Influenza, Human; D013058:Mass Spectrometry / Q000379:methods*; D029121:Metapneumovirus; D012136:Respiratory Syncytial Viruses; D012141:Respiratory Tract Infections / Q000175:diagnosis* / Q000821:virology; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods*; D013997:Time Factors; D014777:Virus Diseases / Q000175:diagnosis* / Q000821:virology; D014780:Viruses / Q000737:chemistry / Q000145:classification* / Q000302:isolation & purification*
keywords: Identification; Influenza virus; LC–MS/MS; MALDI-TOF MS; Mass spectrometry; Virological diagnosis