Simultaneous Dual-Gene Diagnosis of SARS-CoV-2 Based on CRISPR/Cas9-Mediated Lateral Flow Assay
Simultaneous Dual-Gene Diagnosis of SARS-CoV-2 Based on CRISPR/Cas9-Mediated Lateral Flow Assay
复制标题
基于CRISPR/Cas9介导的侧向层析检测的SARS-CoV-2同时双基因诊断
DOI：
10.1002/anie.202014506
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发表时间：
2021-01-26
影响因子：
16.6
通讯作者：
Zhou, Xiaoming
中科院分区：
文献类型：
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作者：
Xiong, Erhu;Jiang, Ling;Zhou, Xiaoming
目前，用于检测SARS-CoV-2的方法很少能够在单一测试中同时检测两个基因，这是提高检测准确性的关键措施，如金标准RT-qPCR方法所采用的。这里开发了CRISPR/Cas9介导的三线侧向流检测（TL-LFA）结合多重逆转录重组酶聚合酶扩增（RT-RPA），用于在单试纸条测试中快速同时检测SARS-CoV-2的双基因。该检测方法的特征在于检测细胞培养的SARS-CoV-2和SARS-CoV-2病毒RNA标准品的包膜（E）和开放阅读框架1ab（Orf 1ab）基因，显示灵敏度为100个RNA拷贝/反应（25 μ L）。此外，64份鼻咽拭子样本的双基因分析显示100%阴性预测一致性和97.14%阳性预测一致性。该平台将为低资源地区的COVID-19或其他传染病的诊断提供更准确和便捷的途径。
Few methods for the detection of SARS-CoV-2 currently have the capability to simultaneously detect two genes in a single test, which is a key measure to improve detection accuracy, as adopted by the gold standard RT-qPCR method. Developed here is a CRISPR/Cas9-mediated triple-line lateral flow assay (TL-LFA) combined with multiplex reverse transcription-recombinase polymerase amplification (RT-RPA) for rapid and simultaneous dual-gene detection of SARS-CoV-2 in a single strip test. This assay is characterized by the detection of envelope (E) and open reading frame 1ab (Orf1ab) genes from cell-cultured SARS-CoV-2 and SARS-CoV-2 viral RNA standards, showing a sensitivity of 100 RNA copies per reaction (25 mu L). Furthermore, dual-gene analysis of 64 nasopharyngeal swab samples showed 100 % negative predictive agreement and 97.14 % positive predictive agreement. This platform will provide a more accurate and convenient pathway for diagnosis of COVID-19 or other infectious diseases in low-resource regions.