4.7 Review

Advances in Field Detection Based on CRISPR/Cas System

Journal

ACS SYNTHETIC BIOLOGY
Volume 10, Issue 11, Pages 2824-2832

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.1c00401

Keywords

Infectious diseases; CRISPR/Cas; nucleic acid detection; POCT; four aspects

Funding

  1. National Science and Technology Specific Projects [2018ZX10301407]
  2. Key Scientific Research Projects in Colleges and Universities of Henan Province [20A330004]
  3. National Natural Science Foundation of China [82073618, 82002147]

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The CRISPR/Cas system provides a rapid and accurate nucleic acid detection method for on-site POCT, with features such as short detection cycles, low cost, and high sensitivity, which are crucial in interrupting outbreaks of infectious diseases.
Rapid and accurate diagnostic methods are essential to interrupt outbreaks of infectious diseases such as COVID-19. However, the most commonly used nucleic acid detection method, qPCR or RT-qPCR, takes several hours to complete and requires highly sophisticated equipment. Recently, an emerging nucleic acid detection method based on the CRISPR/Cas system has reduced the reliance on qPCR. It has several important features that make it suitable for on-site POCT (point-of-care testing), including short detection cycles, low cost, high sensitivity, and the ability to be combined with different readout methods. This review briefly introduces the steps of CRISPR/Cas detection and then summarizes the current advances of CRISPR/Cas-based POCT from four steps: nucleic acid extraction, target amplification, CRISPR/Cas-based signal generation, and signal output. Finally, we discuss the advantages and challenges of CRISPR-based POCT and describe the future research perspectives for CRISPR.

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