Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system.
Instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using self-contained microfluidic system.
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DOI：
10.1016/j.bios.2021.113865
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发表时间：
2022-03-01
影响因子：
12.6
通讯作者：
Liu C
中科院分区：
文献类型：
--
作者：
Li Z;Ding X;Yin K;Avery L;Ballesteros E;Liu C
快速灵敏地检测严重急性呼吸综合征冠状病毒2型（SARS-CoV-2）对于2019冠状病毒病（COVID-19）的早期诊断和及时治疗至关重要。然而，目前的检测方法通常依赖于昂贵且笨重的仪器。在这里，我们开发了一种简单，灵敏，无仪器，基于CRISPR的SARS-CoV-2诊断方法，使用自包含的微流体系统。微流控芯片在一个封闭的微流控平台中集成了等温扩增、CRISPR切割和侧向流检测，实现了无污染的可视化检测。为了简化设备的操作和运输，我们将CRISPR试剂冻干在反应室中，并将液体溶液预先储存在泡罩中。我们采用了一种低成本的便携式暖手器来孵育微流控芯片，而不需要电力。这个独立的微流控系统可以检测到100个SARS-CoV-2 RNA拷贝。此外，我们通过检测24份COVID-19临床鼻咽拭子样本对我们的方法进行了临床验证，实现了出色的灵敏度（94.1%），特异性（100%）和准确性（95.8%）。这种简单、灵敏、价格实惠的微流体系统代表了COVID-19和其他传染病的即时诊断的一种有前途的工具。
Rapid and sensitive detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is critical for early diagnostics and timely medical treatment of coronavirus disease 2019 (COVID-19). However, current detection methods typically rely on expensive and bulky instrumentation. Here, we developed a simple, sensitive, instrument-free, CRISPR-based diagnostics of SARS-CoV-2 using a self-contained microfluidic system. The microfluidic chip integrates isothermal amplification, CRISPR cleavage, and lateral flow detection in a single, closed microfluidic platform, enabling contamination-free, visual detection. To simplify the operation and transportation of the device, we lyophilized the CRISPR reagents in the reaction chamber and pre-stored the liquid solutions in blisters. We employed a low-cost, portable hand warmer to incubate the microfluidic chip without the need for electricity. The self-contained microfluidic system can detect down to 100 copies of SARS-CoV-2 RNA. Further, we clinically validated our method by detecting 24 COVID-19 clinical nasopharyngeal swab samples, achieving excellent sensitivity (94.1%), specificity (100%), and accuracy (95.8%). This simple, sensitive, and affordable microfluidic system represents a promising tool for point-of-care diagnostics of COVID-19 and other infectious diseases.
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DOI：
10.1073/pnas.2010254117
发表时间：
2020-11-24
影响因子：
11.1
作者：
Ramachandran A;Huyke DA;Sharma E;Sahoo MK;Huang C;Banaei N;Pinsky BA;Santiago JG
通讯作者：
Santiago JG
DOI：
10.1016/j.medj.2020.12.010
发表时间：
2021-03-12
期刊：
Med (New York, N.Y.)
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通讯作者：
Grubaugh ND
影响因子：
64.5
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通讯作者：
Ott M
DOI：
10.1016/j.snb.2016.01.073
发表时间：
2016-06-28
期刊：
Sensors and actuators. B, Chemical
影响因子：
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作者：
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通讯作者：
Liu C
影响因子：
7.4
作者：
Rey EG;O'Dell D;Mehta S;Erickson D
通讯作者：
Erickson D