Dual-CRISPR/Cas12a-Assisted RT-RAA for Ultrasensitive SARS-CoV-2 Detection on Automated Centrifugal Microfluidics

Chen, Yong1,2; Zong, Nan1,2; Ye, Feidi3; Mei, Yixin1; Qu, Jiuxin3; Jiang, Xingyu1,2

2022-07-01

10.1021/acs.analchem.2c00638

ANALYTICAL CHEMISTRY   Impact Factor & Quartile

0003-2700

1520-6882

The clustered regularly interspaced short palindromic repeats (CRISPR)-based nucleic acid detection can be combined with recombinase-aided amplification (RAA) to enable rapid, accurate, and early detection of SARS-CoV-2. Current CRISPR-based approaches to detecting viral nucleic acid typically require immense manual operations to transfer RPA amplicons for CRISPR detection or suffer from compromised sensitivity by mixing the competing RPA amplification and CRISPR detection. Here, we develop dual-CRISPR/Cas12a-assisted RT-RAA assay and a "sample-to-answer " centrifugal microfluidic platform that can automatically detect 1 copy/mu L of the SARS-CoV-2 within 30 min. This chip separates the amplification (RAA) from detection (CRISPR), such that sensitivity is maximized and the time consumption is decreased by a factor of 3. For the 26 positive and 8 negative clinical SARS-CoV-2 samples, this automated centrifugal microfluidics achieved 100% accuracy compared to the gold standard RT-PCR technique. This point-of-care test, with the advantages of being one-step, automated, rapid, and sensitive, will have a significant potential for clinical diagnosis and disease prevention.

SCI ; EI

English

NI Journal Papers

First ; Corresponding

National Key R&D Program of China[2018YFA0902600] ; National Natural Science Foundation of China["22104048","21535001","81730051","1761142006"] ; Guangdong Province["2022B1212010003","2019ZT08Y191","2020A1515010586"] ; Department of Education of Guangdong Province[2019KQNCX130] ; Shenzhen Key Laboratory of Smart Healthcare Engineering[ZDSYS20200811144003009] ; Tencent Foundation["2018YFA0902600","22104048","21535001"] ; [KQTD20190929172743294] ; [JCYJ20200109141231365] ; [JCYJ20190809144005609]

Chemistry

Chemistry, Analytical

WOS:000826302200001

AMER CHEMICAL SOC

20223012418006

Automation ; Centrifugation ; Coronavirus ; Microfluidics ; Polymerase chain reaction

Medicine and Pharmacology:461.6 ; Microfluidics:632.5.1 ; Automatic Control Principles and Applications:731 ; Biochemistry:801.2 ; Chemical Operations:802.3

CHEMISTRY

Web of Science

1.Southern Univ Sci & Technol, Dept Biomed Engn, Shenzhen Key Lab Smart Healthcare Engn, Shenzhen 518055, Guangdong, Peoples R China
2.Southern Univ Sci & Technol, Guangdong Prov Key Lab Adv Biomat, Shenzhen 518055, Guangdong, Peoples R China
3.Southern Univ Sci & Technol, Shenzhen Peoples Hosp 3, Dept Clin Lab, Natl Clin Res Ctr Infect Dis,Hosp 2, Shenzhen 518114, Guangdong, Peoples R China

Chen, Yong,Zong, Nan,Ye, Feidi,et al. Dual-CRISPR/Cas12a-Assisted RT-RAA for Ultrasensitive SARS-CoV-2 Detection on Automated Centrifugal Microfluidics[J]. ANALYTICAL CHEMISTRY,2022,94:9603-9609.

Chen, Yong,Zong, Nan,Ye, Feidi,Mei, Yixin,Qu, Jiuxin,&Jiang, Xingyu.(2022).Dual-CRISPR/Cas12a-Assisted RT-RAA for Ultrasensitive SARS-CoV-2 Detection on Automated Centrifugal Microfluidics.ANALYTICAL CHEMISTRY,94,9603-9609.

Chen, Yong,et al."Dual-CRISPR/Cas12a-Assisted RT-RAA for Ultrasensitive SARS-CoV-2 Detection on Automated Centrifugal Microfluidics".ANALYTICAL CHEMISTRY 94(2022):9603-9609.