Development and evaluation of a rapid CRISPR-based diagnostic for COVID-19

Tieying Hou; Weiqi Zeng; Minling Yang; Wenjing Chen; Lili Ren; Jingwen Ai; Ji Wu; Yalong Liao; Xuejing Gou; Yongjun Li; Xiaorui Wang; Hang Su; Bing Gu; Jianwei Wang; Teng Xu

doi:10.1371/journal.ppat.1008705

Development and evaluation of a rapid CRISPR-based diagnostic for COVID-19

PLoS Pathog. 2020 Aug 27;16(8):e1008705. doi: 10.1371/journal.ppat.1008705. eCollection 2020 Aug.

Authors

Tieying Hou¹, Weiqi Zeng^{2

3}, Minling Yang², Wenjing Chen², Lili Ren⁴, Jingwen Ai⁵, Ji Wu¹, Yalong Liao¹, Xuejing Gou², Yongjun Li², Xiaorui Wang², Hang Su², Bing Gu^{6

7}, Jianwei Wang⁴, Teng Xu^{2

3}

Affiliations

¹ Laboratory Medicine, Provincial People's Hospital, Guangdong Academy of Medical Sciences Guangzhou, Guangdong, China.
² Vision Medicals Center for Infectious Diseases, Guangdong, China.
³ Key Laboratory of Animal Gene Editing and Animal Cloning in Yunnan Province and College of Veterinary Medicine, Yunnan Agricultural University, Kunming, China.
⁴ NHC Key Laboratory of Systems Biology of Pathogens and Christophe Mérieux Laboratory, IPB, CAMS-Fondation Mérieux, Institute of Pathogen Biology (IPB), Chinese Academy of Medical Sciences (CAMS) & Peking Union Medical College, Beijing, China.
⁵ Department of Infectious Diseases, Huashan Hospital affiliated to Fudan University, Shanghai, China.
⁶ Medical Technology School of Xuzhou Medical University, Xuzhou, China.
⁷ Department of Laboratory Medicine, Affiliated Hospital of Xuzhou Medical University, Xuzhou, China.

Abstract

The recent outbreak of human infections caused by SARS-CoV-2, the third zoonotic coronavirus has raised great public health concern globally. Rapid and accurate diagnosis of this novel pathogen posts great challenges not only clinically but also technologically. Metagenomic next-generation sequencing (mNGS) and reverse-transcription PCR (RT-PCR) have been the most commonly used molecular methodologies. However, each has their own limitations. In this study, we developed an isothermal, CRISPR-based diagnostic for COVID-19 with near single-copy sensitivity. The diagnostic performances of all three technology platforms were also compared. Our study aimed to provide more insights into the molecular detection of SARS-CoV-2, and also to present a novel diagnostic option for this new emerging virus.

Publication types

Comparative Study
Research Support, Non-U.S. Gov't

MeSH terms

Bacteria / genetics
Betacoronavirus / genetics*
COVID-19
COVID-19 Testing
CRISPR-Cas Systems / genetics*
Clinical Laboratory Techniques*
Clustered Regularly Interspaced Short Palindromic Repeats / genetics
Coronavirus Infections / diagnosis*
Coronavirus Infections / genetics*
Genes, Viral / genetics
Genome, Viral / genetics
High-Throughput Nucleotide Sequencing / methods
Humans
Molecular Diagnostic Techniques / economics
Molecular Diagnostic Techniques / methods
Nucleic Acid Amplification Techniques / economics
Nucleic Acid Amplification Techniques / methods
Pandemics
Pneumonia, Viral / diagnosis*
Pneumonia, Viral / genetics*
Reverse Transcriptase Polymerase Chain Reaction / methods
SARS-CoV-2
Sensitivity and Specificity

Supplementary concepts

LAMP assay

Grants and funding

This work was supported by the "Peak Project" Scientific Research Special Funding [DFJH2020019], the National Major Science & Technology Project for Control and Prevention of Major Infectious Diseases in China [2017ZX10103004], the Guangdong Science and Technology Department [2018B020241002] and the Guangzhou Science, Technology and Innovation Commission [201803010094]. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.