Progress of Clustered Regularly Interspaced Short Palindromic Repeats-associated Proteins Technology in Point-of-Care Testing for Disease

Disease prevention and control have always been crucial to public health and national security. Point-of-care testing (POCT), a method that utilizes portable instruments and reagents to obtain rapid test results at the sampling site, has demonstrated significant advantages and development potential in disease prevention and control while effectively improving diagnostic efficiency. However, many of the reported POCT methods have been limited in practical use due to their low sensitivity. In recent years, the clustered regularly interspaced short palindromic repeats (CRISPR)-CRISPR associated proteins (CRISPR/Cas) system, with its powerful trans-cleavage activity, has gained prominence as an effective signal amplification strategy in detection fields. The integration of CRISPR/Cas systems with POCT has led to rapid advancements in detecting both nucleic acid and non-nucleic acid biomarkers. This paper outlined CRISPR/Cas system types, sensing principles, and target analytes, and categorized CRISPR-based POCT detection methods by signal output modes, including fluorescence, electrochemistry, solution colorimetry, test strip colorimetry, and Raman spectroscopy. Finally, it summarized current developments in this field while discussing existing challenges and future directions.