上海交通大学王鹏飞团队报道了用于核酸敏感检测的可编程DNAzyme。相关研究成果发表在2024年1月30日出版的《德国应用化学》。

体液中的核酸是用于疾病分子诊断的新兴生物标志物，但由于缺乏灵敏的检测方法，其临床应用受到阻碍。

该文中，研究人员报道了一种灵敏的核酸检测方法SPOT（用于核酸检测的灵敏环启动的DNAzyme生物传感器）的开发，该方法将具有内切核酸酶能力的催化DNAzyme设计成统一的单链变构生物传感器。一旦特定序列的核酸靶标与其变构模块结合，SPOT就会被激活，从而能够连续切割分子报告子。SPOT为低丰度核酸的灵敏、方便和成本效益检测提供了一个高度强大的平台。

为了进行临床验证，研究人员证明SPOT可以检测血清miRNA，用于诊断乳腺癌、胃癌和前列腺癌。此外，SPOT对来自临床拭子的严重急性呼吸系统综合征冠状病毒2型RNA表现出强大的检测性能，具有高灵敏度和特异性。最后，SPOT与侧流分析等即时检测模式兼容。因此，该SPOT可以作为一种强大的检测方法，对各种核酸靶点进行灵敏检测，从而在临床中进行分子诊断。

附：英文原文

Title: A Programmable DNAzyme for the Sensitive Detection of Nucleic Acids

Author: Chenzhi Shi, Donglei Yang, Xiaowei Ma, Li Pan, Yuanchuan Shao, Gaurav Arya, Yonggang Ke, Chuan Zhang, Fuan Wang, Xiaolei Zuo, Min Li, Pengfei Wang

Issue&Volume: 2024-01-30

Abstract: Nucleic acids in biofluids are emerging biomarkers for the molecular diagnostics of diseases, but their clinical use has been hindered by the lack of sensitive detection assays. Herein, we report the development of a sensitive nucleic acid detection assay named SPOT (sensitive loop-initiated DNAzyme biosensor for nucleic acid detection) by rationally designing a catalytic DNAzyme of endonuclease capability into a unified one-stranded allosteric biosensor. SPOT is activated once a nucleic acid target of a specific sequence binds to its allosteric module to enable continuous cleavage of molecular reporters. SPOT provides a highly robust platform for sensitive, convenient and cost-effective detection of low-abundance nucleic acids. For clinical validation, we demonstrated that SPOT could detect serum miRNAs for the diagnostics of breast cancer, gastric cancer and prostate cancer. Furthermore, SPOT exhibits potent detection performance over SARS-CoV-2 RNA from clinical swabs with high sensitivity and specificity. Finally, SPOT is compatible with point-of-care testing modalities such as lateral flow assays. Hence, we envision that SPOT may serve as a robust assay for the sensitive detection of a variety of nucleic acid targets enabling molecular diagnostics in clinics.

DOI: 10.1002/anie.202320179

Source: https://onlinelibrary.wiley.com/doi/10.1002/anie.202320179