RT-RPA-Cas12a-based assay facilitates the discrimination of SARS-CoV-2 variants of concern

Tang，Guiyue1; Zhang，Zilong2; Tan，Wei1; Long，Fei1; Sun，Jingxian1; Li，Yingying1; Zou，Siwei1; Yang，Yujiao3; Cai，Kezhu1,4; Li，Shenwei2; Wang，Zhiyi2; Liu，Jiakun1; Mao，Guobing1; Ma，Yingxin1; Zhao，Guo Ping1,3,5; Tian，Zhen Gan2; Zhao，Wei1

2023-04-15

10.1016/j.snb.2023.133433

SENSORS AND ACTUATORS B-CHEMICAL   影响因子和分区

0925-4005

0925-4005

Timely and accurate detection of SARS-CoV-2 variants of concern (VOCs) is urgently needed for pandemic surveillance and control. Great efforts have been made from a mass of scientists in increasing the detection sensitivity and operability, and reducing the turn-around time and cost. Here, we report a nucleic acid testing-based method aiming to detect and discriminate SARS-CoV-2 mutations by combining RT-RPA and CRISPR-Cas12a detecting assays (RRCd). With a detection limit of 10 copies RNA/reaction, RRCd was validated in 194 clinical samples, showing 89% positive predictive agreement and 100% negative predictive agreement, respectively. Critically, using specific crRNAs, representatives of single nucleotide polymorphisms and small deletions in SARS-CoV-2 VOCs including N501Y, T478K and ΔH69-V70 were discriminated by RRCd, demonstrating 100% specificity in clinical samples with C < 33. The method completes within 65 min and could offer visible results without using any electrical devices, which probably facilitate point-of-care testing of SARS-CoV-2 variants and other epidemic viruses.

CRISPR Point-of-care testing RRCd SARS-CoV-2 Variant of concern

SCI ; EI

英语

其他

National Key Research and Development Program of China["2019YFA0904003","2020YFA0909100"] ; Strategic Priority Research Program of the Chinese Academy of Sciences, China[XDB38020300] ; Guangdong Basic and Applied Basic Research Foundation[2021A1515012511] ; General Administration of Customs Project[2020HK003]

Chemistry ; Electrochemistry ; Instruments & Instrumentation

Chemistry, Analytical ; Electrochemistry ; Instruments & Instrumentation

WOS:000982531800001

ELSEVIER SCIENCE SA

20230613540549

Diseases ; Nucleic acids

Biological Materials and Tissue Engineering:461.2 ; Medicine and Pharmacology:461.6 ; Biochemistry:801.2 ; Organic Compounds:804.1

CHEMISTRY

2-s2.0-85147224318

Scopus

1.CAS Key Laboratory of Quantitative Engineering Biology,Shenzhen Institute of Synthetic Biology,Shenzhen Institutes of Advanced Technology,Chinese Academy of Sciences,Shenzhen,518055,China
2.Shanghai International Travel Healthcare Center,Shanghai,Shanghai Customs District P. R. China,200335,China
3.CAS Key Laboratory of Synthetic Biology,CAS Center for Excellence in Molecular Plant Sciences,Chinese Academy of Sciences,Shanghai,200032,China
4.School of Engineering,Southern University of Science and Technology,Shenzhen,518055,China
5.Department of Microbiology and Microbial Engineering,School of Life Sciences,Fudan University,Shanghai,200433,China

Tang，Guiyue,Zhang，Zilong,Tan，Wei,et al. RT-RPA-Cas12a-based assay facilitates the discrimination of SARS-CoV-2 variants of concern[J]. SENSORS AND ACTUATORS B-CHEMICAL,2023,381.

Tang，Guiyue.,Zhang，Zilong.,Tan，Wei.,Long，Fei.,Sun，Jingxian.,...&Zhao，Wei.(2023).RT-RPA-Cas12a-based assay facilitates the discrimination of SARS-CoV-2 variants of concern.SENSORS AND ACTUATORS B-CHEMICAL,381.

Tang，Guiyue,et al."RT-RPA-Cas12a-based assay facilitates the discrimination of SARS-CoV-2 variants of concern".SENSORS AND ACTUATORS B-CHEMICAL 381(2023).