Department of Radiology, Tongji Hospital, Shanghai Frontiers Science Center of Nanocatalytic Medicine, The Institute for Biomedical Engineering &Nano Science, School of Medicine, Tongji University, Shanghai 200065, China.
Galactophore Department, Jingzhou Hospital Affiliated to Yangtze University, Jingzhou 434200, China.
ACS Appl Mater Interfaces. 2024 Aug 28;16(34):44399-44408. doi: 10.1021/acsami.4c05388. Epub 2024 Aug 15.
Nucleic acid detection plays a pivotal role in the accurate diagnosis of diseases. The CRISPR/Cas detection system, noted for its significant utility in a variety of applications, often necessitates enhanced sensitivity or specific signal amplification strategies, particularly for detecting low-abundance biomarkers. In this study, we present a quantum-dot-encoded beads (QDB)-energized CRISPR/Cas12-based lateral-flow assay (QDB-CRISPR-LFA). This method enables amplification-free, sensitive, and rapid detection (<40 min) of BRCA-1. We validated our method using contrived reference samples and nucleic acids extracted from tumor cells. The QDB-CRISPR-LFA provides a visual, more rapid alternative to the traditional BRCA-1 real-time RT-PCR assay. Significantly, through the integration of CRISPR's specificity and the high signal output of QDB, the detection threshold for BRCA-1 has been reduced to the femtomolar level, representing an enhancement of 2-4 orders of magnitude over existing CRISPR/Cas detection methods. This advancement underscores the potential of our approach in advancing nucleic acid detection techniques, which is crucial for the early and precise diagnosis of diseases.
核酸检测在疾病的准确诊断中起着关键作用。CRISPR/Cas 检测系统因其在各种应用中的重要实用性而备受关注，但通常需要增强灵敏度或特定的信号放大策略，特别是用于检测低丰度生物标志物。在这项研究中，我们提出了一种量子点编码珠（QDB）激发的基于 CRISPR/Cas12 的侧流分析（QDB-CRISPR-LFA）。该方法能够实现无扩增、灵敏和快速检测（<40 分钟）BRCA-1。我们使用合成参考样本和从肿瘤细胞中提取的核酸验证了我们的方法。QDB-CRISPR-LFA 为传统的 BRCA-1 实时 RT-PCR 检测提供了一种更快速的可视化替代方法。重要的是，通过整合 CRISPR 的特异性和 QDB 的高信号输出，BRCA-1 的检测阈值已降低到飞摩尔水平，比现有 CRISPR/Cas 检测方法提高了 2-4 个数量级。这一进展突显了我们的方法在推进核酸检测技术方面的潜力，这对于疾病的早期和精确诊断至关重要。
