Detection of SARS-CoV-2 with Solid-State CRISPR-Cas12a-Assisted Nanopores
Detection of SARS-CoV-2 with Solid-State CRISPR-Cas12a-Assisted Nanopores
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DOI：
10.1021/acs.nanolett.1c02974
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发表时间：
2021-09-20
期刊：
影响因子：
10.8
通讯作者：
Guan, Weihua
中科院分区：
文献类型：
--
作者：
Nouri, Reza;Jiang, Yuqian;Guan, Weihua
SARS-CoV-2的爆发导致COVID-19疾病在全球蔓延。对SARS-CoV-2的特异性和敏感性检测有助于早期干预，防止疾病传播。在这里，我们提出了一种固态CRISPR-Cas 12 a辅助纳米孔（SCAN）传感策略，用于SARS-CoV-2的特异性检测。我们引入了一种纳米孔大小的计数方法来测量报告分子的切割率，该切割率用作阳性/阴性分类的标准。开发并验证了动力学切割模型以预测报告分子大小分布。该模型揭示了SARS-CoV-2扫描的灵敏度、周转时间和假阳性率之间的权衡。通过预扩增和30分钟的CRISPR Cas 12 a测定，我们实现了对其他常见人类冠状病毒的优异特异性，并且在95%的置信水平下，病毒RNA的检测限为13.5拷贝/μ L（22.5 aM）。结果表明，该方法可用于SARS-CoV-2的快速、灵敏和特异性检测。
The outbreak of the SARS-CoV-2 caused the disease COVID-19 to spread globally. Specific and sensitive detection of SARS-CoV-2 facilitates early intervention and prevents the disease from spreading. Here, we present a solid-state CRISPR-Cas12a-assisted nanopore (SCAN) sensing strategy for the specific detection of SARS-CoV-2. We introduced a nanopore-sized counting method to measure the cleavage ratio of reporters, which is used as a criterion for positive/negative classification. A kinetic cleavage model was developed and validated to predict the reporter size distributions. The model revealed the trade-offs between sensitivity, turnaround time, and false-positive rate of the SARS-CoV-2 SCAN. With preamplification and a 30 min CRISPR Cas12a assay, we achieved excellent specificity against other common human coronaviruses and a limit of detection of 13.5 copies/mu L (22.5 aM) of viral RNA at a confidence level of 95%. These results suggested that the SCAN could provide a rapid, sensitive, and specific analysis of SARS-CoV-2.