A Rapid DIgital Crispr Approach (RADICA) for the detection and absolute quantification of nucleic acids

Xiaolin Wu; Joshua K. Tay; Chuan Keng Goh; Cheryl Chan; Yie Hou Lee; Stacy L. Springs; De Yun Wang; Kwok Seng Loh; Timothy K. Lu; Hanry Yu

doi:10.1117/12.2614195

4 March 2022 A Rapid DIgital Crispr Approach (RADICA) for the detection and absolute quantification of nucleic acids

Xiaolin Wu, Joshua K. Tay, Chuan Keng Goh, Cheryl Chan, Yie Hou Lee, Stacy L. Springs, De Yun Wang, Kwok Seng Loh, Timothy K. Lu, Hanry Yu

Author Affiliations +

Proceedings Volume PC11979, Frontiers in Biological Detection: From Nanosensors to Systems XIV; PC1197905 (2022) https://doi.org/10.1117/12.2614195
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

Rapid diagnostics of adventitious agents in biopharmaceutical/cell manufacturing release testing and the fight against viral infection have become critical. Quantitative real-time PCR and CRISPR-based methods rapidly detect DNA/RNA in 1 h but suffer from inter-site variability. Absolute quantification of DNA/RNA by methods such as digital PCR reduce this variability but are currently too slow for wider application. Here, we report a RApid DIgital Crispr Approach (RADICA) for absolute quantification of nucleic acids in 40-60 min. Using SARS-CoV-2 and Epstein-Barr virus (EBV) as a proof-of-concept target, RADICA allows for absolute quantification with high accuracy and low variability, no cross-reactivity to similar targets, and high tolerance to human background DNA. RADICA therefore enables rapid and sensitive absolute quantification of nucleic acids which can be widely applied across clinical, research, and biomanufacturing areas.

Conference Presentation

Citation Download Citation

Xiaolin Wu, Joshua K. Tay, Chuan Keng Goh, Cheryl Chan, Yie Hou Lee, Stacy L. Springs, De Yun Wang, Kwok Seng Loh, Timothy K. Lu, and Hanry Yu "A Rapid DIgital Crispr Approach (RADICA) for the detection and absolute quantification of nucleic acids", Proc. SPIE PC11979, Frontiers in Biological Detection: From Nanosensors to Systems XIV, PC1197905 (4 March 2022); https://doi.org/10.1117/12.2614195

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