A multiplex method for detection of SARS-CoV-2 variants based on MALDI-TOF mass spectrometry

IF 3.5 Q1 PUBLIC, ENVIRONMENTAL & OCCUPATIONAL HEALTH

Biosafety and Health Pub Date : 2023-04-01 DOI:10.1016/j.bsheal.2023.02.003

Ziyuan Zhao , Liying Sun , Liqin Wang , Xiaodong Li , Junping Peng

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引用次数: 1

Abstract

The recent outbreak of the coronavirus disease 2019 (COVID-19) pandemic and the continuous evolution of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have highlighted the significance of new detection methods for global monitoring and prevention. Although quantitative reverse transcription PCR (RT-qPCR), the current gold standard for diagnosis, performs excellently in genetic testing, its multiplexing capability is limited because of the signal crosstalk of various fluorophores. Herein, we present a highly efficient platform which combines 17-plex assays with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), enabling the targeting of 14 different mutation sites of the spike gene. Diagnosis using a set of 324 nasopharyngeal swabs or sputum clinical samples with SARS-CoV-2 MS method was identical to that with the RT-qPCR. The detection consistency of mutation sites was 97.9% (47/48) compared to Sanger sequencing without cross-reaction with other respiratory-related pathogens. Therefore, the MS method is highly potent to track and assess SARS-CoV-2 changes in a timely manner, thereby aiding the continuous response to viral variation and prevention of further transmission.

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基于MALDI-TOF质谱法的多重检测SARS-CoV-2变异体方法

最近爆发的2019冠状病毒病（新冠肺炎）大流行和严重急性呼吸综合征冠状病毒2（SARS-CoV-2）的持续演变突出了新检测方法对全球监测和预防的重要性。尽管目前的诊断金标准定量逆转录聚合酶链式反应（RT-qPCR）在基因检测中表现出色，但由于各种荧光团的信号串扰，其多路复用能力受到限制。在此，我们提出了一种高效的平台，它将17丛分析与基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF MS）相结合，能够靶向刺突基因的14个不同突变位点。使用一组324份鼻咽拭子或痰临床样本进行诊断的严重急性呼吸系统综合征冠状病毒2型MS方法与RT-qPCR方法相同。与Sanger测序相比，突变位点的检测一致性为97.9%（47/48），没有与其他呼吸道相关病原体发生交叉反应。因此，MS方法在及时跟踪和评估严重急性呼吸系统综合征冠状病毒2型的变化方面非常有效，从而有助于对病毒变异的持续反应和预防进一步传播。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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