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Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay

Author

Listed:
  • Xiong Ding

    (University of Connecticut Health Center)

  • Kun Yin

    (University of Connecticut Health Center)

  • Ziyue Li

    (University of Connecticut Health Center)

  • Rajesh V. Lalla

    (University of Connecticut Health Center)

  • Enrique Ballesteros

    (University of Connecticut Health Center)

  • Maroun M. Sfeir

    (University of Connecticut Health Center)

  • Changchun Liu

    (University of Connecticut Health Center)

Abstract

The recent outbreak of novel coronavirus (SARS-CoV-2) causing COVID-19 disease spreads rapidly in the world. Rapid and early detection of SARS-CoV-2 facilitates early intervention and prevents the disease spread. Here, we present an All-In-One Dual CRISPR-Cas12a (AIOD-CRISPR) assay for one-pot, ultrasensitive, and visual SARS-CoV-2 detection. By targeting SARS-CoV-2’s nucleoprotein gene, two CRISPR RNAs without protospacer adjacent motif (PAM) site limitation are introduced to develop the AIOD-CRISPR assay and detect the nucleic acids with a sensitivity of few copies. We validate the assay by using COVID-19 clinical swab samples and obtain consistent results with RT-PCR assay. Furthermore, a low-cost hand warmer (~$0.3) is used as an incubator of the AIOD-CRISPR assay to detect clinical samples within 20 min, enabling an instrument-free, visual SARS-CoV-2 detection at the point of care. Thus, our method has the significant potential to provide a rapid, sensitive, one-pot point-of-care assay for SARS-CoV-2.

Suggested Citation

  • Xiong Ding & Kun Yin & Ziyue Li & Rajesh V. Lalla & Enrique Ballesteros & Maroun M. Sfeir & Changchun Liu, 2020. "Ultrasensitive and visual detection of SARS-CoV-2 using all-in-one dual CRISPR-Cas12a assay," Nature Communications, Nature, vol. 11(1), pages 1-10, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-18575-6
    DOI: 10.1038/s41467-020-18575-6
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    Citations

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    Cited by:

    1. Jiongyu Zhang & Chengyu Hou & Changchun Liu, 2024. "CRISPR-powered quantitative keyword search engine in DNA data storage," Nature Communications, Nature, vol. 15(1), pages 1-12, December.
    2. Margot Karlikow & Evan Amalfitano & Xiaolong Yang & Jennifer Doucet & Abigail Chapman & Peivand Sadat Mousavi & Paige Homme & Polina Sutyrina & Winston Chan & Sofia Lemak & Alexander F. Yakunin & Adam, 2023. "CRISPR-induced DNA reorganization for multiplexed nucleic acid detection," Nature Communications, Nature, vol. 14(1), pages 1-11, December.
    3. Yunxiang Wang & Hong Chen & Kai Lin & Yongjun Han & Zhixia Gu & Hongjuan Wei & Kai Mu & Dongfeng Wang & Liyan Liu & Ronghua Jin & Rui Song & Zhen Rong & Shengqi Wang, 2024. "Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device," Nature Communications, Nature, vol. 15(1), pages 1-11, December.
    4. Zhichen Xu & Dongjuan Chen & Tao Li & Jiayu Yan & Jiang Zhu & Ting He & Rui Hu & Ying Li & Yunhuang Yang & Maili Liu, 2022. "Microfluidic space coding for multiplexed nucleic acid detection via CRISPR-Cas12a and recombinase polymerase amplification," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    5. Jeong Moon & Changchun Liu, 2023. "Asymmetric CRISPR enabling cascade signal amplification for nucleic acid detection by competitive crRNA," Nature Communications, Nature, vol. 14(1), pages 1-11, December.

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