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

Dan Xiong,Qianyun Li,Junwei Ye,Wenjun Dai,Guanghui Tang,Yingzhen Jiao,Jiaqiang Pan,Jiaojiao Gong,Chen Chen,Huiling Huang,Wei Wu,Xiuming Zhang,Guande Li,Liang Xue,Nansong Liu,Huina Deng,Xuanxuan Zhang,Bill Sugden

doi:10.1371/journal.pbio.3000978

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.

Highlights

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which emerged in the city of Wuhan, China, has become a serious public health concern [1]
The existing nucleic acid detection methods, such as quantitative reverse transcription polymerase chain reaction (RT-quantitative polymerase chain reaction (qPCR)), are sensitive and accurate, but inadequate access to reagents and machinery has limited their application for disease control
We present an approach for the visual detection of the ORF1ab and N genes of SARS-CoV-2 using clustered regularly interspaced short palindromic repeats (CRISPR)-Cas12a system

Summary

Introduction

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which emerged in the city of Wuhan, China, has become a serious public health concern [1]. Rapid detection of SARS-CoV-2 with CRISPR-Cas12a study design, data collection and analysis, decision to publish, or preparation of the manuscript. We present an approach for the visual detection of the ORF1ab and N genes of SARS-CoV-2 using CRISPR-Cas12a system.

Results

Conclusion