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Simultaneous Detection of SARS-CoV-2 and Six Other Human Coronaviruses by Multiplex PCR and MALDI-TOF MS
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  • Tingting Liu,
  • Lin Kang,
  • Yanwei Li,
  • Jing Huang,
  • Zishuo Guo,
  • Yi Hu,
  • Zhixiang Zhai,
  • XIaoping Kang,
  • Tao Jiang,
  • Hao Li,
  • Hexing Song,
  • Jing Wang,
  • Shan Gao,
  • Jiaxin Li,
  • Xiaoguang Zhou,
  • Yuan Yuan,
  • Jinglin Wang,
  • Baohua Zhao,
  • Wenwen Xin
Tingting Liu
Hebei Normal University
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Lin Kang
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Yanwei Li
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Jing Huang
Hebei Normal University
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Zishuo Guo
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Yi Hu
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Zhixiang Zhai
Rongzhi Biological Technology (Qingdao) Co., Ltd.
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XIaoping Kang
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Tao Jiang
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Hao Li
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Hexing Song
Rongzhi Biological Technology (Qingdao) Co., Ltd.
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Jing Wang
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Shan Gao
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Jiaxin Li
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Xiaoguang Zhou
Rongzhi Biological Technology (Qingdao) Co., Ltd.
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Yuan Yuan
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Jinglin Wang
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology
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Baohua Zhao
Hebei Normal University
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Wenwen Xin
Academy of Military Medical Sciences Institute of Microbiology and Epidemiology

Corresponding Author:xinww@hotmail.com

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Abstract

The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is challenging the health systems worldwide, and large population testing is a vital step to control this pandemic. Here, we developed a new method (named HCoV-MS), which combines multiplex PCR with matrix-assisted laser desorption/ionization-time of flight mass spectrometry to simultaneously detect and differentiate seven human coronaviruses (HCoVs). The HCoV-MS method had good specificity and sensitivity, with a detection limit of 1-5 copies/reaction. To validate the HCoV-MS method, we tested 151 clinical samples, and the results showed good concordance with real-time PCR. In addition, 41 D614G variants were identified, which were consistent with the sequencing results. This method was also used in EQAE-SARS-COV in 2020, and all the samples were accurately identified. Taken together, HCoV-MS could be used as an effective method for large-scale detection. It was also capable of detecting key single nucleotide polymorphism about variants.