Rapid and generic identification of influenza A and other respiratory viruses with mass spectrometry.
Rapid and generic identification of influenza A and other respiratory viruses with mass spectrometry.
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DOI：
10.1016/j.jviromet.2014.11.014
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发表时间：
2015-03
影响因子：
3.1
通讯作者：
Paauw A
中科院分区：
文献类型：
--
作者：
Majchrzykiewicz-Koehorst JA;Heikens E;Trip H;Hulst AG;de Jong AL;Viveen MC;Sedee NJ;van der Plas J;Coenjaerts FE;Paauw A
开发了一种制备用于质谱鉴定的病毒的方法。样品制备方法快速、简单且稳定。这使得能够对培养的呼吸道病毒进行通用且灵敏的识别。同时鉴定混合病毒培养物也是可能的。快速识别现有和新出现的呼吸道病毒对于抗击疫情和流行病至关重要。基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）是细菌诊断中快速可靠的鉴定方法，但尚未用于病毒学诊断。质谱系统已被研究用于呼吸道病毒的鉴定。然而，样品制备方法既费力又费时。在这项研究中，开发了一种可靠且快速的样品制备方法，可以鉴定培养的呼吸道病毒。用所开发的样品制备方法处理十倍连续稀释的十种甲型流感病毒株培养物、甲型流感病毒与人偏肺病毒或呼吸道合胞病毒的混合样品以及重构的临床样品。随后，对肽进行 MALDI-TOF MS 和液相色谱串联质谱 (LC-MS/MS) 分析。使用 MALDI-TOF MS 在 3 小时内对甲型流感病毒株进行亚型鉴定，使用 LC-MS/MS 在 6 小时内鉴定出甲型流感病毒株（不包括培养时间）。与 MALDI-TOF MS 相比，LC-MS/MS 的灵敏度更高。此外，LC-MS/MS 能够区分混合样品中的两种病毒，并能够从重构的临床样品中识别病毒。改进且快速的样品制备方法的开发使得能够通过质谱法对培养的呼吸道病毒进行通用且快速的鉴定。
A method to prepare viruses for identification by mass spectrometry was developed. The sample preparation method was rapid, simple and robust. That allowed for generic and sensitive identification of cultured respiratory viruses. Also simultaneous identification of mixed viral cultures was possible. 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.
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DOI：
10.1016/s0140-6736(10)61459-6
发表时间：
2011-04-09
期刊：
Lancet (London, England)
影响因子：
--
作者：
Ruuskanen O;Lahti E;Jennings LC;Murdoch DR
通讯作者：
Murdoch DR
影响因子：
9.4
作者：
van Veen, S. Q.;Claas, E. C. J.;Kuijper, Ed J.
通讯作者：
Kuijper, Ed J.
影响因子：
9.4
作者：
Fernandes, Neil D.;Downard, Kevin M.
通讯作者：
Downard, Kevin M.
影响因子：
11.8
作者：
Seng, Piseth;Drancourt, Michel;Raoult, Didier
通讯作者：
Raoult, Didier
影响因子：
4.2
作者：
Nguyen, An P.;Downard, Kevin M.
通讯作者：
Downard, Kevin M.