Advances in One-Tube Biomarker Detection Technology Based on CRISPR/Cas Signal Amplification: A Review

The clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated proteins (Cas) system and isothermal amplification methods combined in one tube can not only shorten the detection time but also greatly decrease the risk of nucleic acid aerosol contamination. However, the single-tube solution resulted in a significant decrease in detection sensitivity due to the incompatibility of the two systems. This review provides a comprehensive analysis of current advancements aimed at resolving these challenges. This review categorizes these advancements into four primary strategies: 1) optimization of the buffer system; 2) spatial isolation within the reaction tube; 3) chemical isolation of reaction components; and 4) amplification-free detection methods. In addition to summarizing the latest progress, it emphasizes the biological research developments related to the one-tube strategy for detecting various markers through CRISPR/Cas signal amplification. This review aims to serve as a valuable reference for researchers exploring this rapidly evolving field.