Amplification-Free Detection of SARS-CoV-2 and Respiratory Syncytial Virus Using CRISPR Cas13a and Graphene Field-Effect Transistors

Huijie Li; Jie Yang; Guangfu Wu; Zhengyan Weng; Yang Song; Yuxuan Zhang; Jeffrey A Vanegas; Lori Avery; Zan Gao; He Sun; Yupeng Chen; Kevin D Dieckhaus; Xue Gao; Yi Zhang

doi:10.1002/anie.202203826

Amplification-Free Detection of SARS-CoV-2 and Respiratory Syncytial Virus Using CRISPR Cas13a and Graphene Field-Effect Transistors

Angew Chem Int Ed Engl. 2022 Aug 8;61(32):e202203826. doi: 10.1002/anie.202203826. Epub 2022 May 31.

Authors

Huijie Li^{1

2}, Jie Yang³, Guangfu Wu^{1

2}, Zhengyan Weng^{1

2}, Yang Song^{1

2}, Yuxuan Zhang^{1

2}, Jeffrey A Vanegas³, Lori Avery⁴, Zan Gao^{1

2}, He Sun^{1

2}, Yupeng Chen¹, Kevin D Dieckhaus⁵, Xue Gao^{3

6

7}, Yi Zhang^{1

2}

Affiliations

¹ Department of Biomedical Engineering, University of Connecticut, Storrs, CT 06269, USA.
² Institute of Materials Science, University of Connecticut, Storrs, CT 06269, USA.
³ Department of Chemical and Biomolecular Engineering, Rice University, Houston, TX 77005, USA.
⁴ Department of Pathology and Laboratory Medicine, UConn Health, Farmington, CT 06030, USA.
⁵ Division of Infectious Diseases, Department of Medicine, UConn Health, Farmington, CT 06030, USA.
⁶ Department of Bioengineering, Rice University, Houston, TX 77005, USA.
⁷ Department of Chemistry, Rice University, Houston, TX 77005, USA.

Abstract

The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) systems have recently received notable attention for their applications in nucleic acid detection. Despite many attempts, the majority of current CRISPR-based biosensors in infectious respiratory disease diagnostic applications still require target preamplifications. This study reports a new biosensor for amplification-free nucleic acid detection via harnessing the trans-cleavage mechanism of Cas13a and ultrasensitive graphene field-effect transistors (gFETs). CRISPR Cas13a-gFET achieves the detection of SARS-CoV-2 and respiratory syncytial virus (RSV) genome down to 1 attomolar without target preamplifications. Additionally, we validate the detection performance using clinical SARS-CoV-2 samples, including those with low viral loads (Ct value >30). Overall, these findings establish our CRISPR Cas13a-gFET among the most sensitive amplification-free nucleic acid diagnostic platforms to date.

Keywords: Amplification-Free Detection; Biosensors; CRISPR Cas13a; Graphene Field-Effect Transistors; SARS-CoV-2.

Publication types

Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

COVID-19*
CRISPR-Cas Systems
Graphite*
Humans
Nucleic Acids*
Respiratory Syncytial Viruses
SARS-CoV-2 / genetics

Substances

Nucleic Acids
Graphite

Grants and funding

R01 AR072027/AR/NIAMS NIH HHS/United States