Proteotyping SARS-CoV-2 Virus from Nasopharyngeal Swabs: A Proof-of-Concept Focused on a 3 Min Mass Spectrometry Window
Proteotyping SARS-CoV-2 Virus from Nasopharyngeal Swabs: A Proof-of-Concept Focused on a 3 Min Mass Spectrometry Window
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DOI：
10.1021/acs.jproteome.0c00535
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发表时间：
2020-11-06
影响因子：
4.4
通讯作者：
Armengaud, Jean
中科院分区：
文献类型：
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作者：
Gouveia, Duarte;Miotello, Guylaine;Armengaud, Jean
对临床样本中的严重急性呼吸综合征冠状病毒2（SARSCoV-2）进行快速但敏感、特异和高通量的检测是诊断感染者和更好地控制病毒传播的关键。无需特定试剂的PCR和免疫诊断替代方法值得研究，不仅用于抗击COVID-19大流行，还用于检测其他紧急病原体威胁。在这里，我们建议使用串联质谱检测SARS-CoV-2标记肽在鼻咽拭子。我们记录了来自这些样品中存在的微生物群的信号很低，并且在解释在肽组景观的受限窗口上获得的鸟枪蛋白质组数据时可以被忽略。在这项概念验证研究中，使用加标不同量纯化SARS-CoV-2病毒材料的类似鼻咽拭子开发了nanoLC-MS/MS采集方法，然后成功应用于COVID-19临床样本。我们认为，来自核衣壳蛋白的肽ADETQALPQR和GFYAQGSR是最感兴趣的，因为它们的信号是强烈的，并且它们的洗脱可以在测试条件下在3分钟的窗口内获得。这些结果为基于质谱的时间有效的病毒诊断测试的开发铺平了道路。
Rapid but yet sensitive, specific, and high-throughput detection of the severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) in clinical samples is key to diagnose infected people and to better control the spread of the virus. Alternative methodologies to PCR and immunodiagnostics that would not require specific reagents are worthy to investigate not only for fighting the COVID-19 pandemic but also to detect other emergent pathogenic threats. Here, we propose the use of tandem mass spectrometry to detect SARS-CoV-2 marker peptides in nasopharyngeal swabs. We documented that the signal from the microbiota present in such samples is low and can be overlooked when interpreting shotgun proteomic data acquired on a restricted window of the peptidome landscape. In this proof-of-concept study, simili nasopharyngeal swabs spiked with different quantities of purified SARS-CoV-2 viral material were used to develop a nanoLC-MS/MS acquisition method, which was then successfully applied on COVID-19 clinical samples. We argue that peptides ADETQALPQR and GFYAQGSR from the nucleocapsid protein are of utmost interest as their signal is intense and their elution can be obtained within a 3 min window in the tested conditions. These results pave the way for the development of time-efficient viral diagnostic tests based on mass spectrometry.