Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device
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-dep...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2024-04-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-47518-8 |
| _version_ | 1827277012202422272 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Yunxiang Wang |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-24T07:13:45Z |
| format | Article |
| id | doaj.art-19447f3357234585b569e7937d67c57e |
| institution | Directory Open Access Journal |
| issn | 2041-1723 |
| language | English |
| last_indexed | 2024-04-24T07:13:45Z |
| publishDate | 2024-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj.art-19447f3357234585b569e7937d67c57e2024-04-21T11:24:26ZengNature PortfolioNature Communications2041-17232024-04-0115111110.1038/s41467-024-47518-8Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic deviceYunxiang Wang0Hong Chen1Kai Lin2Yongjun Han3Zhixia Gu4Hongjuan Wei5Kai Mu6Dongfeng Wang7Liyan Liu8Ronghua Jin9Rui Song10Zhen Rong11Shengqi Wang12Bioinformatics Center of AMMSBioinformatics Center of AMMSDepartment of Clinical Laboratory, Air Force Medical Center, Air Force Medical UniversityBioinformatics Center of AMMSBeijing Ditan Hospital, Capital Medical UniversityBioinformatics Center of AMMSBioinformatics Center of AMMSBioinformatics Center of AMMSBioinformatics Center of AMMSBeijing Ditan Hospital, Capital Medical UniversityBeijing Ditan Hospital, Capital Medical UniversityBioinformatics Center of AMMSBioinformatics Center of AMMSAbstract 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.https://doi.org/10.1038/s41467-024-47518-8 |
| spellingShingle | 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 Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device Nature Communications |
| title | Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device |
| title_full | Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device |
| title_fullStr | Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device |
| title_full_unstemmed | Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device |
| title_short | Ultrasensitive single-step CRISPR detection of monkeypox virus in minutes with a vest-pocket diagnostic device |
| title_sort | ultrasensitive single step crispr detection of monkeypox virus in minutes with a vest pocket diagnostic device |
| url | https://doi.org/10.1038/s41467-024-47518-8 |
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