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

• Published in: Nature communications Vol. 15; no. 1; p. 3279
• Main Authors: Wang, Yunxiang, Chen, Hong, Lin, Kai, Han, Yongjun, Gu, Zhixia, Wei, Hongjuan, Mu, Kai, Wang, Dongfeng, Liu, Liyan, Jin, Ronghua, Song, Rui, Rong, Zhen, Wang, Shengqi
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 16.04.2024; Nature Publishing Group; Nature Portfolio
• Subjects: 45/71; 631/1647/350/59; 631/208/4041; 631/326/2521; 692/699/255/2514; 692/700/139/1512; Biological Assay; Cell Death; CRISPR; CRISPR-Cas Systems; Exanthema; Global health; Humanities and Social Sciences; Humans; Lysis; Male; Monkeypox; Monkeypox virus; multidisciplinary; Nucleic Acid Amplification Techniques; Nucleic acids; Nucleotide sequence; Nucleotidyltransferases; Public health; Recombinase; Recombinases; Saliva; Science; Science (multidisciplinary); Sensitivity and Specificity; Viruses
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-024-47518-8

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. The recent monkeypox outbreak highlighted the need for rapid and accurate diagnosis of this disease. Here, authors develop an ultrasensitive and streamlined CRISPR assay using miniaturized device, which can detect monkeypox virus in rash fluid swab, oral swab, saliva, and urine within 15 minutes.