DNA糖基化酶的单分子检测及其在癌症早期诊断中的应用研究

Maintaining genomic DNA stability is a necessity for all organisms’ survival. DNA glycosylase is an important initiating enzyme of cellular base excision repair pathway, which is responsible for the repair of various DNA lesions and the maintenance of genomic integrity. The dysregulation of DNA glycosylase activity is associated with a variety of human diseases including cancers. Thus, the development of ultrasensitive techniques for DNA glycosylase is of significance to early diagnosis and treatment of cancers. Single-molecule detection has remarkable advantages of high signal-to-noise ratio, high sensitivity and low sample consumption, and therefore has been widely applied to the clinical research. We will develop single-molecule detection-based techniques for the detection of DNA glycosylase with rapidity, accuracy and sensitivity, and further explore their application to complex biological samples and intact cells. We hope the single-molecule detection-based techniques will provide a powerful tool for biomedical research and clinical medical diagnosis.

基因组DNA的稳定性是所有生物体赖以生存的必要条件。DNA糖基化酶是细胞内碱基切除修复途径的重要起始酶，负责修复多种DNA损伤并维持基因组DNA的完整性。DNA糖基化酶的异常将导致包括癌症在内的许多疾病的发生，因此开发DNA糖基化酶超灵敏检测技术对疾病的早期诊断和治疗具有十分重大的意义。单分子检测技术具有信噪比高、灵敏度高、样品所需量少等优点，被广泛应用于生物医学研究。申请人拟开发快速、准确、超灵敏检测DNA糖基化酶的单分子检测技术，并探索其在复杂样品体系、完整细胞内的应用，为生物医学研究和临床医学诊断提供强有力的工具。

本项目针对与生物标志物 DNA 糖基化酶的超灵敏检测，开展了一系列国际前沿性的研究工作。从特定核苷酸探针设计、检测灵敏度提高以及复杂环境应用等角度出发，我们将核苷酸扩增技术、荧光共振能量转移技术和化学发光技术与单分子检测技术结合起来，创建了一整套单分子检测理论体系，搭建了一种简单、灵敏、特异的 DNA 糖基化酶检测平台，开发了一系列多功能的纳米光学探针与高灵敏光学检测技术，实现了对重大疾病生物标志物的准确、快速、高灵敏的检测。项目实施中的核心科研成果主要由 SCI 论文体现，在国际顶级期刊《Chemical Science》、《Analytical Chemistry》、《Biosensors and Bioelectronics》等上共计发表高水平SCI 论文 11 篇，累积影响因子达到 91.024，超额完成目标，相关的核心技术已申请国家发明专利共计 11 项。