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Simultaneous Detection of Seven Human Coronaviruses by Multiplex PCR and MALDI-TOF MS
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Tingting Liu, Lin Kang, Yanwei Li, Jing Huang, Zishuo Guo, Jinglin Xu, Yi Hu, Zhixiang Zhai, Xiaoping Kang, Tao Jiang, Hao Li, Hexing Song, Jing Wang, Shan Gao, Jiaxin Li, Xiaoguang Zhou, Yuan Yuan, Baohua Zhao, Jinglin Wang and Wenwen Xin
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Abstract
Human coronaviruses (HCoVs) are associated with a range of respiratory symptoms. The discovery of severe acute respiratory syndrome (SARS)-CoV, Middle East respiratory syndrome, and SARS-CoV-2 pose a significant threat to human health. In this study, we developed a method (HCoV-MS) that combines multiplex
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Human coronaviruses (HCoVs) are associated with a range of respiratory symptoms. The discovery of severe acute respiratory syndrome (SARS)-CoV, Middle East respiratory syndrome, and SARS-CoV-2 pose a significant threat to human health. In this study, we developed a method (HCoV-MS) that combines multiplex PCR with matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS), to detect and differentiate seven HCoVs simultaneously. The HCoV-MS method had high specificity and sensitivity, with a 1–5 copies/reaction detection limit. To validate the HCoV-MS method, we tested 163 clinical samples, and the results showed good concordance with real-time PCR. Additionally, the detection sensitivity of HCoV-MS and real-time PCR was comparable. The HCoV-MS method is a sensitive assay, requiring only 1 μL of a sample. Moreover, it is a high-throughput method, allowing 384 samples to be processed simultaneously in 30 min. We propose that this method be used to complement real-time PCR for large-scale screening studies.
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