Recent advances in integrated CRISPR/Cas biosensing for aquatic food safety: overcoming the matrix interference and the difficulties in point-of-care testing

Rapid, sensitive, and on-site detection of pathogenic bacteria in aquatic-product and aquaculture-associated samples is critical for food safety. Although CRISPR/Cas has emerged as a powerful biosensing tool, its practical application is hindered by complex aquatic matrices and the difficulty of integrating pretreatment, amplification, and signal readout into point-of-care testing (POCT) workflows. This review examines integrated CRISPR/Cas biosensing strategies for pathogenic bacterial detection in aquatic matrices. It outlines the enzymatic features of core effectors, reviews pretreatment methods for complex samples, and summarizes how CRISPR/Cas is combined with isothermal amplification to improve analytical sensitivity. It also compares biosensing platforms from the perspective of matrix interference and field applicability. The analysis indicates that isothermal amplification is essential for detecting low-abundance targets in complex aquatic matrices. Among current strategies, ratiometric fluorescence, magnetic separation-assisted sensing, and signal-on electrochemical reporting show particular promise because they improve calibration, reduce matrix background, and limit nonspecific signal loss, respectively. Future progress depends on standardized sample processing, quantitative multiplexing, and automated "sample-in, result-out" systems for real-world deployment.