Sensitive and Easy-Read CRISPR Strip for COVID-19 Rapid Point-of-Care Testing
Sensitive and Easy-Read CRISPR Strip for COVID-19 Rapid Point-of-Care Testing
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用于 COVID-19 快速护理点测试的灵敏且易于读取的 CRISPR 条带
DOI：
10.1089/crispr.2020.0138
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发表时间：
2021-06-01
期刊：
影响因子：
3.7
通讯作者：
Sun, Yansong
中科院分区：
文献类型：
--
作者：
Li, Hao;Dong, Xue;Sun, Yansong
对严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）的快速和临床敏感检测在接触者追踪和遏制COVID-19大流行方面发挥着重要作用。最近开发的一种可现场部署的具有侧向流条带的成簇规则间隔短回文重复序列（CRISPR）检测方法显示出对SARS-CoV-2的即时检测的希望。然而，基于试纸条的检测方法的检测限（10-100拷贝/μ L）远低于基于荧光的检测方法。在这项研究中，我们开发了一种易于读取和灵敏度增强（ERASE）的条带，以可视化CRISPR检测的结果，并将灵敏度提高到1拷贝/μ L，并具有明确的易于读取的结果。使用从中国患者中收集的盲样的649份临床样本，我们验证了我们用于SARS-CoV-2 RNA检测的ERASE检测方法，阳性预测一致率为90.67%，阴性预测一致率为99.21%。总之，我们的研究提供了一种定制的CRISPR条带，用于SARS-CoV-2检测的简单，快速，超灵敏和高度特异性的检测。
Rapid and clinically sensitive detection of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) play an important role in the contact tracing and containment of the COVID-19 pandemic. A recently developed field-deployable clustered regularly interspaced short palindromic repeats (CRISPR) detection assay with lateral flow strips shows promise for point-of-care detection of SARS-CoV-2. However, the limit of detection of paper strip-based assays (10-100 copies/mu L) is much lower than that of fluorescence-based detection methods. In this study, we developed an easy-readout and sensitive enhanced (ERASE) strip to visualize the results of CRISPR detection and improve the sensitivity to 1 copy/mu L with an unambiguous easy-read result. Using 649 clinical samples from blind specimens collected from patients in China, we validated our ERASE assay for SARS-CoV-2 RNA detection with 90.67% positive predictive agreement and 99.21% negative predictive agreement. In conclusion, our study provided a customized CRISPR strip for use in a simple, rapid, ultrasensitive, and highly specific assay for SARS-CoV-2 detection.