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Research Article

Nanomechanical assay for ultrasensitive and rapid detection of SARS-CoV-2 based on peptide nucleic acid

Yu Wang1, Tianhao Yan1, Kainan Mei1, Depeng Rao1, Wenjie Wu1, Ye Chen1, Yongpei Peng1, Jianye Wang2, Shangquan Wu1( ), Qingchuan Zhang1( )
CAS Key Laboratory of Mechanical Behavior and Design of Material, Department of Modern Mechanics, University of Science and Technology of China, Hefei 230027, China
Reproductive Medicine Center, Department of Obstetrics and Gynecology, First Affiliated Hospital of Anhui Medical University, Hefei 230022, China
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Graphical Abstract

Herein, a nanomechanical sensoring assay based on peptide nucleic acid and microcantilever array has been developed for ultrasensitive and amplification-free detection of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2). Thanks to the stress enhancement brought by PNA and the mechanics-based microcantilever which is very sensitive to stress, the method has an extremely low detection limit of 50 copies/mL of the N gene of SARS-CoV-2 within 1 h.

Abstract

The massive global spread of the COVID-19 pandemic makes the development of more effective and easily popularized assays critical. Here, we developed an ultrasensitive nanomechanical method based on microcantilever array and peptide nucleic acid (PNA) for the detection of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) RNA. The method has an extremely low detection limit of 0.1 fM (105 copies/mL) for N-gene specific sequence (20 bp). Interestingly, it was further found that the detection limit of N gene (pharyngeal swab sample) was even lower, reaching 50 copies/mL. The large size of the N gene dramatically enhances the sensitivity of the nanomechanical sensor by up to three orders of magnitude. The detection limit of this amplification-free assay method is an order of magnitude lower than RT-PCR (500 copies/mL) that requires amplification. The non-specific signal in the assay is eliminated by the in-situ comparison of the array, reducing the false-positive misdiagnosis rate. The method is amplification-free and label-free, allowing for accurate diagnosis within 1 h. The strong specificity and ultra-sensitivity allow single base mutations in viruses to be distinguished even at very low concentrations. Also, the method remains sensitive to fM magnitude lung cancer marker (miRNA-155). Therefore, this ultrasensitive, amplification-free and inexpensive assay is expected to be used for the early diagnosis of COVID-19 patients and to be extended as a broad detection tool.

Keywords

SARS-CoV-2nucleic acid assaynanomechanical sensoringpeptide nucleic acidamplification-free

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Nano Research
Volume 16 Issue 1,
January 2023
Pages 1183-1195
DOI: 10.1007/s12274-022-4333-3
Cite this article:
Wang Y, Yan T, Mei K, et al. Nanomechanical assay for ultrasensitive and rapid detection of SARS-CoV-2 based on peptide nucleic acid. Nano Research, 2023, 16(1): 1183-1195. https://doi.org/10.1007/s12274-022-4333-3
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Received: 09 February 2022
Revised: 14 March 2022
Accepted: 16 March 2022
Published: 19 May 2022
© Tsinghua University Press 2022
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