Detection of SARS-CoV-2 with Solid-State CRISPR-Cas12a-Assisted Nanopores,Nano Letters

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Detection of SARS-CoV-2 with Solid-State CRISPR-Cas12a-Assisted Nanopores
Nano Letters ( IF 9.6 ) Pub Date : 2021-09-20 , DOI: 10.1021/acs.nanolett.1c02974
Reza Nouri ₁ , Yuqian Jiang _{2,

3} , Zifan Tang ₁ , Xiaojun Lance Lian _{2,

3,

4} , Weihua Guan _{1,

2}

Affiliation

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/μL (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.

中文翻译：

使用固态 CRISPR-Cas12a 辅助纳米孔检测 SARS-CoV-2

SARS-CoV-2 的爆发导致新冠肺炎 (COVID-19) 在全球范围内传播。对 SARS-CoV-2 进行特异性和灵敏的检测有助于早期干预并防止疾病传播。在这里，我们提出了一种固态 CRISPR-Cas12a 辅助纳米孔 (SCAN) 传感策略，用于特异性检测 SARS-CoV-2。我们引入了纳米孔大小的计数方法来测量报告基因的裂解率，将其用作阳性/阴性分类的标准。开发并验证了动力学裂解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 CRISPR Cas12a 测定，我们对其他常见人类冠状病毒实现了出色的特异性，病毒 RNA 的检测限为 13.5 拷贝/μL (22.5 aM)，置信度为 95%。这些结果表明 SCAN 可以对 SARS-CoV-2 提供快速、灵敏和特异性的分析。

更新日期：2021-10-13

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