Quantum-Dot-Encoded\nBeads-Enhanced CRISPR/Cas-Based\nLateral-Flow Assay for the Amplification-Free, Sensitive, and Rapid\nDetection of Nucleic Acids in Breast Cancer

Nucleic\nacid detection plays a pivotal role in the accurate diagnosis\nof diseases. The CRISPR/Cas detection system, noted for its significant\nutility in a variety of applications, often necessitates enhanced\nsensitivity or specific signal amplification strategies, particularly\nfor detecting low-abundance biomarkers. In this study, we present\na quantum-dot-encoded beads (QDB)-energized CRISPR/Cas12-based lateral-flow\nassay (QDB-CRISPR-LFA). This method enables amplification-free, sensitive,\nand rapid detection (<40 min) of BRCA-1. We validated our method\nusing contrived reference samples and nucleic acids extracted from\ntumor cells. The QDB-CRISPR-LFA provides a visual, more rapid alternative\nto the traditional BRCA-1 real-time RT-PCR assay. Significantly, through\nthe integration of CRISPR’s specificity and the high signal\noutput of QDB, the detection threshold for BRCA-1 has been reduced\nto the femtomolar level, representing an enhancement of 2–4\norders of magnitude over existing CRISPR/Cas detection methods. This\nadvancement underscores the potential of our approach in advancing\nnucleic acid detection techniques, which is crucial for the early\nand precise diagnosis of diseases.