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

Yunxiang Wang, Hong Chen, Kai Lin, Yongjun Han, Zhixia Gu, Hongjuan Wei, Kai Mu, Dongfeng Wang, Liyan Liu, Ronghua Jin (), Rui Song (), Zhen Rong () and Shengqi Wang ()
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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

Nature Communications, 2024, vol. 15, issue 1, 1-11

Abstract: 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.

Date: 2024
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DOI: 10.1038/s41467-024-47518-8

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