Multiplexed discrimination of SARS-CoV-2 variants via duplex-specific nuclease combined MALDI-TOF MS.

Autor: Han G; Department of Chemistry, and Shanghai Stomatological Hospital, Fudan University, Shanghai, 200000, China.; Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, Hangzhou, 310022, Zhejiang, China., Deng W; College of Pharmaceutical Sciences, Zhejiang University, Hangzhou, 310058, Zhejiang, China., Lyu Q; Bioyong Technologics Inc, Beijing, 100176, China., Ma Q; Bioyong Technologics Inc, Beijing, 100176, China., Qiao L; Department of Chemistry, and Shanghai Stomatological Hospital, Fudan University, Shanghai, 200000, China. liang_qiao@fudan.edu.cn.
Jazyk: angličtina
Zdroj: Analytical and bioanalytical chemistry [Anal Bioanal Chem] 2024 Mar; Vol. 416 (8), pp. 1833-1842. Date of Electronic Publication: 2024 Feb 17.
DOI: 10.1007/s00216-024-05202-2
Abstrakt: The frequent mutations in SARS-CoV-2 significantly increase the virus's pathogenicity and transmissibility while also diminishing the effectiveness of vaccines. Consequently, assays capable of rapidly and simultaneously identifying multiple SARS-CoV-2 variants are essential for large-scale applications that aim to monitor the evolution of the virus. In this work, we propose a method combining duplex-specific nuclease (DSN)-assisted cyclic amplification with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) detection, enabling the simultaneous identification of multiple SARS-CoV-2 variants at high-throughput. Due to the high specificity of DSN, single-base mutations can be resolved by the method. With ultra-sensitive detection by MALDI-TOF MS, a limit of detection of 100 pM viral RNA fragment was demonstrated. The assay was used for simultaneous identification and typing of SARS-CoV-2 Alpha, Beta, and Delta variants. The whole assay can be accomplished within 3 h, and the amplification is performed under constant temperature, making the technique simple in operation and efficient. It is also feasible to extend the technique to the detection of many other variants of the virus. We expect that the method can add value to the rapid screening of viral variants and can play an important role in pandemic control.
(© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)
Databáze: MEDLINE
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