基于等温扩增的脱氧核酶荧光生物传感器检测肿瘤标志物技术的研究

Accurate, sensitive, and rapid detection of tumor markers has great clinical significance in the cancer diagnosis and treatment. However, accurate detection of biomarkers in clinical samples is the great challenging because of its complex composition and low concentration. Moreover, a wide range of tumor markers, including DNA, microRNA, proteins and small molecules are detected with requirement of specific methods in clinical diagnosis, which greatly increases the experimental cost and complexity. To overcome the low sensitivity, poor specificity and high cost in the tumor markers detection, we propose to develop a simple, cost-effective, sensitive and specific isothermal exponential amplification-based DNAzyme fluorescence biosensing platform for the detection of tumor markers. This method can convert the different tumor markers to the same DNAzyme sequence by amplification reaction, which decreases the experimental cost and complexity. The isothermal exponential amplification-based DNAzyme fluorescence biosensing platform has the potential to be further applied for the detection of various diseases in the early clinical diagnosis.

准确，灵敏，快速的检测肿瘤标志物在肿瘤的诊断和治疗中具有极其重要的意义。然而，由于临床样本成分复杂和生物标志物浓度偏低，精确的检测临床样本中具有诊断意义的生物标志物面临着巨大挑战。肿瘤标志物的种类繁多，包括DNA，microRNA，蛋白及小分子物质等等，目前在临床检测中针对不同的肿瘤标志物，需要特异的检测方法，因此大大增加了检测的成本费用和复杂性。针对目前临床样品检测灵敏度低、特异性差和费用高的问题，本项目拟发展一种基于等温扩增的脱氧核酶（DNAzyme）荧光检测技术，这种方法通过引入扩增技术将不同的肿瘤标志物通过扩增反应转化成同样的DNAzyme序列，极大减少了检测成本和检测复杂性。实现能够在临床中超灵敏，高特异，简单，低成本的检测各种肿瘤标志物。

本项目发展了基于等温扩增技术和荧光相结合的检测技术，可应用于肿瘤相关生物标志物的超灵敏检测和临床疾病的早期诊断，极大减少了检测成本和检测复杂性。本项目检测了各种肿瘤生物标志物，包括DNA糖基化酶、DNA甲基化转移酶、转录因子、DNA甲基化及DNA损伤等。项目实施中的核心科研成果主要由 SCI 论文体现，我们在国际杂志《Analytical Chemistry》和《Chemical Communications》等发表高水平 SCI 论文14 篇，累积影响因子达到88.511。相关的核心技术已申请国家发明专利5项，授权1项。