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Vol. 14 (2022)

CRISPR-Cas12a-Empowered Electrochemical Biosensor for Rapid and Ultrasensitive Detection of SARS-CoV-2 Delta Variant

https://doi.org/10.1007/s40820-022-00888-4
Chenshuo Wu
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Zhi Chen
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Chaozhou Li
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Yabin Hao
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Yuxuan Tang
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Yuxuan Yuan
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Luxiao Chai
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Taojian Fan
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Jiangtian Yu
Shenzhen International Institute for Biomedical Research, Shenzhen 518116, People’s Republic of China
Xiaopeng Ma
Department of Respiratory, Shenzhen Children’s Hospital, Shenzhen 518038, People’s Republic of China
Omar A. Al‑Hartomy
Department of Physics, Faculty of Science, King Abdulaziz University, Jeddah 21589, Saudi Arabia
S. Wageh
Department of Physics, Faculty of Science, King Abdulaziz University, Jeddah 21589, Saudi Arabia
Abdullah G. Al‑Sehemi
Research Center for Advanced Materials Science (RCAMS), King Khalid University, P.O. Box 9004, Abha 61413, Saudi Arabia
Zhiguang Luo
Zhongmin (Shenzhen) Intelligent Ecology Co., Ltd, Shenzhen 518055, People’s Republic of China
Yaqing He
Shenzhen Center for Disease Control and Prevention, Shenzhen 518055, People’s Republic of China
Jingfeng Li
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China
Zhongjian Xie
Shenzhen International Institute for Biomedical Research, Shenzhen 518116, People’s Republic of China
Han Zhang
International Collaborative Laboratory of 2D, Materials for Optoelectronics Science and Technology of Ministry of Education, Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, Shenzhen University, Shenzhen 518060, People’s Republic of China

Corresponding Author: Han Zhang

hzhang@szu.edu.cn

Nano-Micro Letters, Vol. 14 (2022), Article Number: 159

Abstract

Coronavirus disease 2019 (COVID-19) is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). The gold standard method for the diagnosis of SARS-CoV-2 depends on quantitative reverse transcription-polymerase chain reaction till now, which is time-consuming and requires expensive instrumentation, and the confirmation of variants relies on further sequencing techniques. Herein, we first proposed a robust technique-methodology of electrochemical CRISPR sensing with the advantages of rapid, highly sensitivity and specificity for the detection of SARS-CoV-2 variant. To enhance the sensing capability, gold electrodes are uniformly decorated with electro-deposited gold nanoparticles. Using DNA template identical to SARS-CoV-2 Delta spike gene sequence as model, our biosensor exhibits excellent analytical detection limit (50 fM) and high linearity (R2 = 0.987) over six orders of magnitude dynamic range from 100 fM to 10 nM without any nucleic-acid-amplification assays. The detection can be completed within 1 h with high stability and specificity which benefits from the CRISPR-Cas system. Furthermore, based on the wireless micro-electrochemical platform, the proposed biosensor reveals promising application ability in point-of-care testing.


Highlights:


1 A robust technique-methodology of electrochemical CRISPR sensing is first proposed for the rapid, highly sensitive and specific detection of SARS-CoV-2 variant without any nucleic-acid-amplification assays.
2 Using the DNA template identical to SARS-CoV-2 Delta spike gene sequence as model, our biosensor exhibited excellent analytical detection limit (50 fM) and high linearity (R2 = 0.987) without any amplification assay.
3 Specific crRNA was designed to match the mutation site on nucleic acid sequence of the SARS-CoV-2 Delta variant, presenting programmability, universality, and scalability for diagnosis of other emerging SARS-CoV-2 variants.

Keywords

SARS-COV-2 variant Methodology of electrochemical CRISPR sensing (MOECS) Gold nanoparticles (AuNPs) Point-of-care testing (POCT)
Wu, Chenshuo, Zhi Chen, Chaozhou Li, Yabin Hao, Yuxuan Tang, Yuxuan Yuan, Luxiao Chai, Taojian Fan, Jiangtian Yu, Xiaopeng Ma, Omar A. Al‑Hartomy, S. Wageh, Abdullah G. Al‑Sehemi, Zhiguang Luo, Yaqing He, Jingfeng Li, Zhongjian Xie, and Han Zhang. 2022. “CRISPR-Cas12a-Empowered Electrochemical Biosensor for Rapid and Ultrasensitive Detection of SARS-CoV-2 Delta Variant”. Nano-Micro Letters 14 (August):159. https://doi.org/10.1007/s40820-022-00888-4.
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