Fu Ruijie, Wang Yanzhong, Qiao Sai, Xu Peng, Xianyu Yunlei, Zhang Jun
Department of Clinical Laboratory of Sir Run Run Shaw Hospital, College of Biosystems Engineering and Food Science, Zhejiang University School of Medicine, Hangzhou 310058, People's Republic of China.
Key Laboratory of Precision Medicine in Diagnosis and Monitoring Research of Zhejiang Province, Sir Run Run Shaw Hospital, Hangzhou 310016, People's Republic of China.
ACS Sens. 2025 Feb 28;10(2):1429-1439. doi: 10.1021/acssensors.4c03461. Epub 2025 Jan 25.
The rapid, simple, and sensitive detection of nucleic acid biomarkers plays a significant role in clinical diagnosis. Herein, we develop a label-free and point-of-care approach for isothermal DNA detection through the trans-cleavage activity of CRISPR-Cas12 and the growth of gold nanomaterials in agarose gel. The presence of the target can activate CRISPR-Cas12a to cleave single-stranded DNA, thus modulating the length and number of DNA sequences that mediate the growth of gold nanoparticles (AuNPs) or gold nanorods (AuNRs). Due to the extraordinary plasmonic property of gold nanomaterials, they present characteristic absorption/color after the growth with unique shapes. The sensing strategy is applied to detect BRCA-1, a biomarker related to breast cancer, with limits of detection of 1.72 pM (AuNP-based) and 2.07 pM (AuNR-based). AuNPs/AuNRs can be immobilized in agarose gels that display different colors in the presence of target DNA sequences. The agarose gel-based test allows for a readout by the naked eye or the RGB value with a smartphone. The approach is isothermal and label-free without any surface modification of nanomaterials, which holds great potential for the detection of nucleic acids in clinical applications.
核酸生物标志物的快速、简单且灵敏的检测在临床诊断中发挥着重要作用。在此，我们通过CRISPR-Cas12的反式切割活性以及金纳米材料在琼脂糖凝胶中的生长，开发了一种用于等温DNA检测的无标记即时检测方法。靶标的存在可激活CRISPR-Cas12a切割单链DNA，从而调节介导金纳米颗粒（AuNP）或金纳米棒（AuNR）生长的DNA序列的长度和数量。由于金纳米材料具有非凡的等离子体特性，它们在生长后呈现出具有独特形状的特征吸收/颜色。该传感策略用于检测与乳腺癌相关的生物标志物BRCA-1，基于AuNP的检测限为1.72 pM，基于AuNR的检测限为2.07 pM。AuNP/AuNR可固定在琼脂糖凝胶中，在存在靶DNA序列时显示不同颜色。基于琼脂糖凝胶的检测可通过肉眼或用智能手机读取RGB值进行读数。该方法是等温且无标记的，无需对纳米材料进行任何表面修饰，在临床应用中的核酸检测方面具有巨大潜力。
