Development of a Parallel Reaction Monitoring Mass Spectrometry Assay for the Detection of SARS-CoV-2 Spike Glycoprotein and Nucleoprotein

作者：Lisa H. Cazares, Raghothama Chaerkady, Shao Huan Samuel Weng, Chelsea C. Boo, Raffaello Cimbro, Hsiang-En Hsu, Sarav Rajan, William F. Dall’Acqua, Lori A. Clarke, Kuishu Ren, Patrick M. McTamney, Nicole Kallewaard-LeLay, Mahboobe Ghaedi, Yasuhiro Ikeda, Sonja Hess · 发表于：Analytical Chemistry · 年份：2020 · DOI：10.1021/acs.analchem.0c02288 · 被引用次数：64 · 研究领域：SARS-CoV-2 detection and testing、SARS-CoV-2 and COVID-19 Research、Biosensors and Analytical Detection

High Resolution Image Download MS PowerPoint Slide There is an urgent need for robust and high-throughput methods for SARS-CoV-2 detection in suspected patient samples to facilitate disease management, surveillance, and control. Although nucleic acid detection methods such as reverse transcription polymerase chain reaction (RT-PCR) are the gold standard, during the current pandemic, the deployment of RT-PCR tests has been extremely slow, and key reagents such as PCR primers and RNA extraction kits are at critical shortages. Rapid point-of-care viral antigen detection methods have been previously employed for the diagnosis of respiratory viruses such as influenza and respiratory syncytial viruses. Therefore, the direct detection of SARS-CoV-2 viral antigens in patient samples could also be used for diagnosis of active infection, and alternative methodologies for specific and sensitive viral protein detection should be explored. Targeted mass spectrometry techniques have enabled the identification and quantitation of a defined subset of proteins/peptides at single amino acid resolution with attomole level sensitivity and high reproducibility. Herein, we report a targeted mass spectrometry assay for the detection of SARS-CoV-2 spike protein and nucleoprotein in a relevant biological matrix. Recombinant full-length spike protein and nucleoprotein were digested and proteotypic peptides were selected for parallel reaction monitoring (PRM) quantitation using a high-resolution Orbitr...