Kollhoff Lydia, Kipping Marc, Rauh Manfred, Ceglarek Uta, Barka Günes, Barka Frederik, Sinz Andrea
Department of Pharmaceutical Chemistry and Bioanalytics, Martin Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, D-06120, Halle (Saale), Germany.
Center for Structural Mass Spectrometry, Martin Luther University Halle-Wittenberg, Kurt-Mothes-Str. 3, D-06120, Halle (Saale), Germany.
Clin Proteomics. 2023 Jul 1;20(1):26. doi: 10.1186/s12014-023-09415-y.
We have developed a rapid and highly specific assay for detecting and monitoring SARS-CoV-2 infections by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). As MALDI-TOF mass spectrometers are available in a clinical setting, our assay has the potential to serve as alternative to the commonly used reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). Sample preparation prior to MALDI-TOF-MS involves the tryptic digestion of SARS-CoV-2 proteins, followed by an enrichment of virus-specific peptides from SARS-CoV-2 nucleoprotein via magnetic antibody beads. Our MALDI-TOF-MS method allows the detection of SARS-CoV-2 nucleoprotein in sample collection medium as low as 8 amol/µl. MALDI-TOF mass spectra are obtained in just a few seconds, which makes our MS-based assay suitable for a high-throughput screening of SARS-CoV-2 in healthcare facilities in addition to PCR. Due to the specific detection of virus peptides, different SARS-CoV-2 variants are readily distinguished from each other. Specifically, we show that our MALDI-TOF-MS assay discriminates SARS-CoV-2 strain B.1.617.2 "delta variant" from all other variants in patients' samples, making our method highly valuable to monitor the emergence of new virus variants.
我们开发了一种通过基质辅助激光解吸/电离飞行时间质谱（MALDI-TOF-MS）检测和监测SARS-CoV-2感染的快速且高度特异的检测方法。由于临床环境中可使用MALDI-TOF质谱仪，我们的检测方法有潜力作为常用的逆转录定量聚合酶链反应（RT-qPCR）的替代方法。MALDI-TOF-MS之前的样品制备包括对SARS-CoV-2蛋白进行胰蛋白酶消化，然后通过磁性抗体珠从SARS-CoV-2核蛋白中富集病毒特异性肽段。我们的MALDI-TOF-MS方法能够检测样品采集介质中低至8 amol/µl的SARS-CoV-2核蛋白。只需几秒钟就能获得MALDI-TOF质谱图，这使得我们基于质谱的检测方法除了PCR之外，还适用于医疗机构对SARS-CoV-2进行高通量筛查。由于对病毒肽段的特异性检测，不同的SARS-CoV-2变异株很容易相互区分。具体而言，我们表明我们的MALDI-TOF-MS检测方法能够在患者样本中区分SARS-CoV-2 B.1.617.2毒株“德尔塔变异株”与所有其他变异株，这使得我们的方法对于监测新病毒变异株的出现具有很高的价值。
