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Rapid and generic identification of influenza A and other respiratory viruses with mass spectrometry

Author: Majchrzykiewicz-Koehorst, J.A. · Heikens, E. · Trip, H. · Hulst, A.G. · Jong, A.L. de · Viveen, M.C. · Sedee, N.J.A. · van der Plas, J. · Coenjaerts, F.E.J. · Paauw, A.
Type:article
Date:2015
Publisher: Elsevier
Source:Journal of Virological Methods, 213, 75-83
Identifier: 521644
doi: doi:10.1016/j.jviromet.2014.11.014
Keywords: Biology · Identification · Influenza virus · LC-MS/MS · MALDI-TOF MS · Mass spectrometry · Virological diagnosis · virus protein · Amino acid sequence · Controlled study · Influenza virus A · Intermethod comparison · Liquid chromatography · Matrix assisted laser desorption ionization time of flight mass spectrometry · Metapneumovirus · Nonhuman · Nucleotide sequence · Process development · Reliability · Respiratory syncytial pneumovirus · Respiratory virus · RNA sequence · Sensitivity analysis · Sequence analysis · Tandem mass spectrometry · Virus identification · Virus strain · Bacteria (microorganisms) · Human metapneumovirus · Influenza A virus · Orthomyxoviridae · Respiratory syncytial virus · Life · CBRN - CBRN Protection · TS - Technical Sciences

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 3. h with MALDI-TOF MS and 6. h 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.