Trends in developing one-pot CRISPR diagnostics strategies

HighlightsCRISPR-based nucleic acid detection assays offer a potent and dependable tool in modern nucleic acid detection, owing to their high specificity, rapidity, sensitivity, ease of use, and broad applicability.The combination of the CRISPR system and nucleic acid amplification technology has transformed the modern biomedical landscape, markedly augmenting the specificity of nucleic acid amplification, thereby advancing the field of precision medicine.The one-pot nucleic acid detection assay based on the CRISPR system realizes the integration of nucleic acid amplification and CRISPR detection into a single reaction tube, which not only simplifies the experimental operations but also significantly reduces the risk of aerosol contamination.AbstractThe integration of nucleic acid amplification (NAA) with the CRISPR detection system has led to significant advancements and opportunities for development in molecular diagnostics. Nevertheless, the incompatibility between CRISPR cleavage and NAA has significantly impeded the commercialization of this technology. Currently, several one-pot detection strategies based on CRISPR systems have been devised to address concerns regarding aerosol contamination risk and operational complexity associated with step-by-step detection as well as the sensitivity limitation of conventional one-pot methods. In this review, we provide a comprehensive introduction and outlook of the various solutions of the one-pot CRISPR assay for practitioners who are committed to developing better CRISPR nucleic acid detection technologies to promote the progress of molecular diagnostics.