Signal Amplification Strategies and Advances in CRISPR/Cas Molecular Diagnostics

Accurate detection of infectious diseases and serious diseases is of great importance to public health. CRISPR/Cas has become a highly sensitive and specific molecular diagnostic tool with its programmable recognition and trans-cleavage activity. This article reviews its classification and mechanisms, nucleic acid detection principle, signal amplification strategy, one-tube detection and clinical translation. It is pointed out that pre-amplification coupling, amplification free enhancement and system optimization can significantly improve the sensitivity. One-tube strategies such as physical isolation, glycerol separation, light control, and suboptimal PAM can alleviate aerosol contamination and problems with amplification compatibility, but multiplexing throughput, sample interference, and POCT integration still constrain clinical translation. This study provides a reference for the optimization of CRISPR molecular diagnosis and “sample-in, result-out” application.