Sun Xudong, Li Xiaotong, Jiang Hao, Shan Yongjie, Zhou Sifeng, Wang Zhenxin
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, P. R. China.
School of Applied Chemistry and Engineering, University of Science and Technology of China, Hefei 230026, P. R. China.
Anal Chem. 2025 Jul 1;97(25):13086-13094. doi: 10.1021/acs.analchem.5c00468. Epub 2025 Jun 13.
Specific glycosylation patterns on exosome surfaces represent novel diagnostic biomarkers for cancer liquid biopsy. Lectins can induce exosome aggregation through multiple bindings with exosomal glycoproteins. In this work, we developed a one-pot lectin-induced recombinase polymerase amplification (RPA) and CRISPR/Cas12a-mediated cleavage assay (LI-RPA-CRISPR/Cas12a) for diagnosing colorectal cancer (CRC) through the interactions of abundant α-fucose residues on CRC cell-derived exosome surfaces with Ulex Europaeus Agglutinin I (UEA-I). The combination of a homemade portable isothermal amplification device, the as-proposed LI-RPA-CRISPR/Cas12a exhibits a wide detection range from 2 × 10 to 1 × 10 extracellular vehicles (EVs) μL with a visual limit of detection (LOD) as low as 1.0 × 10 EVs μL, and has been successfully utilized to dynamically monitor the progression of tumors in mice-bearing SW480 CRC subtype at an early stage. After integration with a long short-term memory (LSTM) deep learning model, the LI-RPA-CRISPR/Cas12a achieves accurate diagnosis of primary colorectal cancer with a drop of blood through a smartphone-based data analysis application, reaching an accuracy of 95% in 100 clinical samples. This rapid, sensitive, and user-friendly approach provides a promising platform for point-of-care testing (POCT) diagnosis of CRC, enabling early detection and monitoring of disease progression through a minimally invasive liquid biopsy.
外泌体表面特定的糖基化模式代表了癌症液体活检的新型诊断生物标志物。凝集素可通过与外泌体糖蛋白的多重结合诱导外泌体聚集。在本研究中，我们开发了一种一锅法凝集素诱导的重组酶聚合酶扩增（RPA）和CRISPR/Cas12a介导的切割检测方法（LI-RPA-CRISPR/Cas12a），用于通过结直肠癌（CRC）细胞来源外泌体表面丰富的α-岩藻糖残基与欧洲荆豆凝集素I（UEA-I）的相互作用来诊断结直肠癌。结合自制的便携式等温扩增装置，所提出的LI-RPA-CRISPR/Cas12a检测范围广泛，从2×10到1×10个细胞外囊泡（EVs）/μL，视觉检测限（LOD）低至1.0×10个EVs/μL，并已成功用于在早期动态监测携带SW480 CRC亚型的小鼠肿瘤进展。与长短期记忆（LSTM）深度学习模型整合后，LI-RPA-CRISPR/Cas12a通过基于智能手机的数据分析应用程序，一滴血即可实现原发性结直肠癌的准确诊断，在100个临床样本中准确率达到95%。这种快速、灵敏且用户友好的方法为结直肠癌的即时检测（POCT）诊断提供了一个有前景的平台，能够通过微创液体活检实现疾病进展的早期检测和监测。
