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Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device

Author

Listed:
  • Yunxiang Wang

    (Bioinformatics Center of AMMS)

  • Hong Chen

    (Bioinformatics Center of AMMS)

  • Kai Lin

    (Air Force Medical Center, Air Force Medical University)

  • Yongjun Han

    (Bioinformatics Center of AMMS)

  • Zhixia Gu

    (Capital Medical University)

  • Hongjuan Wei

    (Bioinformatics Center of AMMS)

  • Kai Mu

    (Bioinformatics Center of AMMS)

  • Dongfeng Wang

    (Bioinformatics Center of AMMS)

  • Liyan Liu

    (Bioinformatics Center of AMMS)

  • Ronghua Jin

    (Capital Medical University)

  • Rui Song

    (Capital Medical University)

  • Zhen Rong

    (Bioinformatics Center of AMMS)

  • Shengqi Wang

    (Bioinformatics Center of AMMS)

Abstract

The emerging monkeypox virus (MPXV) has raised global health concern, thereby highlighting the need for rapid, sensitive, and easy-to-use diagnostics. Here, we develop a single-step CRISPR-based diagnostic platform, termed SCOPE (Streamlined CRISPR On Pod Evaluation platform), for field-deployable ultrasensitive detection of MPXV in resource-limited settings. The viral nucleic acids are rapidly released from the rash fluid swab, oral swab, saliva, and urine samples in 2 min via a streamlined viral lysis protocol, followed by a 10-min single-step recombinase polymerase amplification (RPA)-CRISPR/Cas13a reaction. A pod-shaped vest-pocket analysis device achieves the whole process for reaction execution, signal acquisition, and result interpretation. SCOPE can detect as low as 0.5 copies/µL (2.5 copies/reaction) of MPXV within 15 min from the sample input to the answer. We validate the developed assay on 102 clinical samples from male patients / volunteers, and the testing results are 100% concordant with the real-time PCR. SCOPE achieves a single-molecular level sensitivity in minutes with a simplified procedure performed on a miniaturized wireless device, which is expected to spur substantial progress to enable the practice application of CRISPR-based diagnostics techniques in a point-of-care setting.

Suggested Citation

  • 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.
    • Handle: RePEc:nat:natcom:v:15:y:2024:i:1:d:10.1038_s41467-024-47518-8
    DOI: 10.1038/s41467-024-47518-8
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    References listed on IDEAS

    as
    1. Alexandra East-Seletsky & Mitchell R. O’Connell & Spencer C. Knight & David Burstein & Jamie H. D. Cate & Robert Tjian & Jennifer A. Doudna, 2016. "Two distinct RNase activities of CRISPR-C2c2 enable guide-RNA processing and RNA detection," Nature, Nature, vol. 538(7624), pages 270-273, October.
    2. 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.
    3. 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.
    4. Max Kozlov, 2022. "Monkeypox goes global: why scientists are on alert," Nature, Nature, vol. 606(7912), pages 15-16, June.
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