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

• 作 者: Ziyuan Zhao Liying Sun Liqin Wang Xiaodong Li Junping Peng
• 机 构: NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, Chinese Academy o
• 出 处: 生物安全与健康 (英文)
• 发表日期: 2023年05期
• 关键词: COVID-19 SARS-CoV-2 Multiplexing capability MALDI-TOF mass spectrometry Viral va

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.