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

被引:27
|
作者
Majchrzykiewicz-Koehorst, Joanna A. [1 ]
Heikens, Esther [2 ]
Trip, Hein [1 ]
Hulst, Albert G. [1 ]
de Jong, Ad L. [1 ]
Viveen, Marco C. [2 ]
Sedee, Norbert J. A. [1 ]
van der Plas, Jan [3 ]
Coenjaerts, Frank E. J. [2 ]
Paauw, Armand [1 ]
机构
[1] TNO, Netherlands Org Appl Sci Res, Dept CBRN Protect, NL-2288 GJ Rijswijk, Netherlands
[2] Univ Med Ctr Utrecht, Dept Med Microbiol, NL-3584 CX Utrecht, Netherlands
[3] Expert Ctr Force Hlth Protect, Div Hlth Care, Support Command, Minist Def, NL-3941 PW Doorn, Netherlands
关键词
Mass spectrometry; MALDI-TOF MS; LC-MS/MS; Influenza virus; Identification; Virological diagnosis; DESORPTION IONIZATION-TIME; PANDEMIC H1N1; VIRAL LOADS; NEURAMINIDASE; MUTATIONS; BACTERIA; PROTEIN;
D O I
10.1016/j.jviromet.2014.11.014
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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. (C) 2014 The Authors. Published by Elsevier B.V.
引用
收藏
页码:75 / 83
页数:9
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