Detection of monkeypox virus based on a convenient and sensitive single-step RPA-CRISPR/Cas12a strategy
The global outbreak of monkeypox virus (MPXV) has highlighted the need for rapid molecular diagnostics techniques. In this study, a single-step recombinase polymerase amplification (RPA)-CRISPR/Cas12a system was developed for rapid and sensitive detection of MPXV. The limit of detection of this assa...
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| Veröffentlicht in: | RSC advances 2024-05, Vol.14 (21), p.14775-14783 |
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| creator | Yu, Tao Rong, Zhen Gu, Zhixia Wei, Hongjuan Wang, Yunxiang Song, Rui Wang, Shengqi Wang, Shumei |
| description | The global outbreak of monkeypox virus (MPXV) has highlighted the need for rapid molecular diagnostics techniques. In this study, a single-step recombinase polymerase amplification (RPA)-CRISPR/Cas12a system was developed for rapid and sensitive detection of MPXV. The limit of detection of this assay was 1 copy per μL of extracted nucleic acids. A heating lysis method was integrated to further simplify the sample processing workflow and shorten the assay time to 40 min from sample to result. The reaction mixture can be lyophilized to improve its accessibility in resource-limited settings. The analysis results of the proposed single-step RPA-CRISPR/Cas12a assay for clinical MPXV positive and negative samples were 100% consistent with standard PCR assay. These results demonstrate the feasibility and efficiency of this method for rapid and accurate MPXV detection in real-world settings, showcasing its potential utility in urgent and practical settings.
A convenient and sensitive single-step RPA-CRISPR/Cas12a assay was developed for point-of-care detection of monkeypox virus in clinical samples. |
| doi_str_mv | 10.1039/d4ra02049a |
| format | Article |
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| source | DOAJ Directory of Open Access Journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; PubMed Central Open Access |
| subjects | Assaying Chemistry Mpox Nucleic acids Workflow |
| title | Detection of monkeypox virus based on a convenient and sensitive single-step RPA-CRISPR/Cas12a strategy |
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