文献类型：期刊文献

英文题名：Integrating PCR-free amplification and synergistic sensing for ultrasensitive and rapid CRISPR/Cas12a-based SARS-CoV-2 antigen detection

作者：Zhao, Xiangxiang[1,2];Wang, Zhengduo[1,2];Yang, Bowen[1,2];Li, Zilong[3];Tong, Yaojun[4,5];Bi, Yuhai[6];Li, Zhenghong[3];Xia, Xuekui[7];Chen, Xiangyin[1,2];Zhang, Lixin[1,2];Wang, Weishan[3];Tan, Gao-Yi[1,2]

机构：[1]East China Univ Sci & Technol ECUST, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]East China Univ Sci & Technol ECUST, Sch Biotechnol, Shanghai 200237, Peoples R China;[3]Chinese Acad Sci, Inst Microbiol, State Key Lab Microbial Resources, Beijing 100101, Peoples R China;[4]Shanghai Jiao Tong Univ, State Key Lab Microbial Metab, Shanghai 200240, Peoples R China;[5]Shanghai Jiao Tong Univ, Sch Life Sci & Biotechnol, Shanghai 200240, Peoples R China;[6]Chinese Acad Sci, Ctr Influenza Res & Early Warning CASCIRE, CAS TWAS Ctr Excellence Emerging Infect Dis CEEID, Inst Microbiol, Beijing 100101, Peoples R China;[7]Qilu Univ Technol, Biol Inst, Key Biosensor Lab Shandong Prov, Shandong Acad Sci, Jinan 250013, Peoples R China

年份：2021

卷号：6

期号：4

起止页码：283

外文期刊名：SYNTHETIC AND SYSTEMS BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000751793700005)】；

基金：This work was supported by the National Key R&D program of China (2020YFA0907800), the National Natural Science Foundation of China (31922002, 31720103901, 31772242 and 31870040), the 111 Project (B18022), the Fundamental Research Funds for the Central Universities [22221818014], the Youth Innovation Promotion Association CAS (Y202027) to W.W and the Open Project Funding of the State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：CRISPR; Cas12a; Aptamer; Antigen detection; SARS-CoV-2; PCR-Free amplification; Synergistic sensing

摘要：Antigen detection provides particularly valuable information for medical diagnoses; however, the current detection methods are less sensitive and accurate than nucleic acid analysis. The combination of CRISPR/Cas12a and aptamers provides a new detection paradigm, but sensitive sensing and stable amplification in antigen detection remain challenging. Here, we present a PCR-free multiple trigger dsDNA tandem-based signal amplification strategy and a de novo designed dual aptamer synergistic sensing strategy. Integration of these two strategies endowed the CRISPR/Cas12a and aptamer-based method with ultra-sensitive, fast, and stable antigen detection. In a demonstration of this method, the limit of detection was at the single virus level (0.17 fM, approximately two copies/mu L) in SARS-CoV-2 antigen nucleocapsid protein analysis of saliva or serum samples. The entire procedure required only 20 min. Given our system's simplicity and modular setup, we believe that it could be adapted reasonably easily for general applications in CRISPR/Cas12a-aptamer-based detection.