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基于 CRISPR-Cas12a 的 SARS-CoV-2 快速检测。

Rapid detection of SARS-CoV-2 with CRISPR-Cas12a.

机构信息

Medical Laboratory of Shenzhen Luohu People's Hospital, Shenzhen, China.

Yaneng Biotech Co. Ltd., Fosun Pharma, Shenzhen, China.

出版信息

PLoS Biol. 2020 Dec 15;18(12):e3000978. doi: 10.1371/journal.pbio.3000978. eCollection 2020 Dec.

Abstract

The recent outbreak of betacoronavirus Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which is responsible for the Coronavirus Disease 2019 (COVID-19) global pandemic, has created great challenges in viral diagnosis. The existing methods for nucleic acid detection are of high sensitivity and specificity, but the need for complex sample manipulation and expensive machinery slow down the disease detection. Thus, there is an urgent demand to develop a rapid, inexpensive, and sensitive diagnostic test to aid point-of-care viral detection for disease monitoring. In this study, we developed a clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated proteins (Cas) 12a-based diagnostic method that allows the results to be visualized by the naked eye. We also introduced a rapid sample processing method, and when combined with recombinase polymerase amplification (RPA), the sample to result can be achieved in 50 minutes with high sensitivity (1-10 copies per reaction). This accurate and portable detection method holds a great potential for COVID-19 control, especially in areas where specialized equipment is not available.

摘要

最近爆发的β冠状病毒严重急性呼吸综合征冠状病毒 2(SARS-CoV-2)引起了 2019 年冠状病毒病(COVID-19)全球大流行,给病毒诊断带来了巨大挑战。现有的核酸检测方法具有高灵敏度和特异性,但需要复杂的样本处理和昂贵的仪器,这减缓了疾病的检测速度。因此,迫切需要开发一种快速、廉价、灵敏的诊断测试方法,以辅助即时护理点的病毒检测,用于疾病监测。在本研究中,我们开发了一种基于成簇规律间隔短回文重复序列(CRISPR)-CRISPR 相关蛋白(Cas)12a 的诊断方法,可通过肉眼观察结果。我们还引入了一种快速的样本处理方法,当与重组酶聚合酶扩增(RPA)结合使用时,该方法可在 50 分钟内实现高灵敏度(每个反应 1-10 个拷贝)的检测。这种准确、便携的检测方法在 COVID-19 控制方面具有很大的潜力,特别是在缺乏专业设备的地区。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0ad4/7737895/a2ba17394f1a/pbio.3000978.g001.jpg

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