A warm-start digital CRISPR/Cas-based method for the quantitative detection of nucleic acids
Name
2021.06.10.21258725v1.full.pdf
Description
Submitted version
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891.63 KB
Format
Adobe PDF
Checksum (MD5)
7282df60642c339a133474369cce695b
Author(s)
Wu, Xiaolin
Chan, Cheryl
Springs, Stacy L
Lee, Yie Hou
Lu, Timothy K
Yu, Hanry
Date Issued
2022
Journal
Analytica Chimica Acta
Publisher
Elsevier BV
Citation
Wu, Xiaolin, Chan, Cheryl, Springs, Stacy L, Lee, Yie Hou, Lu, Timothy K et al. 2022. "A warm-start digital CRISPR/Cas-based method for the quantitative detection of nucleic acids." Analytica Chimica Acta, 1196.
Version
Original manuscript
Abstract
Nucleic acids-based molecular diagnostic tools incorporating the CRISPR/Cas system are being developed as rapid and sensitive methods for pathogen detection. However, most CRISPR/Cas-based diagnostics lack quantitative detection ability. Here, we report Warm-Start RApid DIgital Crispr Approach (WS-RADICA) for the rapid, sensitive, and quantitative detection of nucleic acids. WS-RADICA detected as little as 1 copy/μl SARS-CoV-2 RNA in 40 min (qualitative detection) or 60 min (quantitative detection). WS-RADICA can be easily adapted to various digital devices: two digital chips were evaluated for both DNA and RNA quantification, with linear dynamic ranges of 0.8-12777 copies/μL for DNA and 1.2-18391 copies/μL for RNA (both R2 values > 0.99). Moreover, WS-RADICA had lower detection limit and higher inhibitor tolerance than a bulk RT-LAMP-Cas12b reaction and similar performance to RT-qPCR and RT-dPCR. To prove its performance on nucleic acids derived from live virus, WS-RADICA was also validated to detect and quantify human adenovirus and herpes simplex virus. Given its speed, sensitivity, quantification capability, and inhibitor tolerance, WS-RADICA shows great promise for a variety of applications requiring nucleic acid quantification.
MIT Department
Massachusetts Institute of Technology. Synthetic Biology Center
Singapore-MIT Alliance in Research and Technology (SMART)
Massachusetts Institute of Technology. Synthetic Biology Center
Massachusetts Institute of Technology. Research Laboratory of Electronics
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Massachusetts Institute of Technology. Department of Biological Engineering
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Creative Commons Attribution-NonCommercial-NoDerivs License
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DOI of Published Version
https://doi.org/10.1016/J.ACA.2022.339494
