Fig. 8

Steps, applications, and advantages of bacterial detection methods based on the CRISPR-Cas system. CRISPR-Cas systems (i.e., Cas9, Cas12, Cas13 and Cas14) have been developed for pathogen diagnosis. First, clinical samples are enriched, and pathogen nucleic acids are extracted and amplified. For RNA samples, reverse-transcription amplification with T7 transcription is required. Then, the target is identified and cleaved by the corresponding CRISPR-Cas system. The sensitivity can be enhanced by fluorescence or lateral flow assays, the results can be observed with the naked eye, and the quantitative detection of pathogens can be realized. In addition, CRISPR-Cas systems can be used for diverse diagnostic purposes, such as distinguishing pathogen genotypes from SNPs, distinguishing ARGs from virulence genes, diagnosing pathogen infections, and performing rapid on-site detection. CRISPR-based pathogen diagnostics facilitate simple portability, high sensitivity and specificity, time savings, and multiple detection. (Created with BioRender.com)