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. 2021 Apr 23;11:639108. doi: 10.3389/fcimb.2021.639108

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Copyright © 2021 Ding, Long, Yuan, Jin, Yang, Chen, Chen and Duan

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Experimental protocol for SARS-CoV-2 detection on CRISPR/Cas detection platform. (1) The extracted SARS-CoV-2 viral RNA is first pre-amplified (RT-RPA and RT-LAMP) into double-stranded DNA (dsDNA). (2) For Cas13a-SHERLOCK assay, dsDNA is first T7 transcribed into single-stranded RNA (ssRNA), followed by activation of the Cas13a cleavage reporter. (3) For Cas12a or Cas12b assays, direct detection of pre-amplified dsDNA enables activation of the Cas12a or Cas12b cleavage reporter. (4) Visualization of CRISPR/Cas assay results by fluorescence and lateral flow readouts.