基于汽凝法的生物芯片的开发与应用研究

Early diagnosis of major diseases (e.g. cancer) is key to reduce mortality. Biodiagnostic technology with low cost, high sensitivity, selectivity, high-throughput and fast detection remains an unmet need for clinical diagnosis, especially in less developed regions. In this project, we innovatively integrate silver staining (<15 min) and vapor condensation-based readout (<1 min) with bio-chips for realizing fast detection. The metallic materials produced by the silver staining can change the wettability and color of the detection points, and convert biological recognition to vapor condensation, forming two-stage amplification, which will greatly improve the detection sensitivity and save time. In addition, we will extend the bio-chips for detection of biomarkers (such as genetic mutations, miRNA, small molecules, ions, proteins, and even cells) involved in biomedical diagnosis. The detection results can be visualized by naked eye, which greatly reduces the cost and well meets the need for application in the resource-limited regions. To meet the need of various detection environment and increase the flexibility of the bio-chips, we will develop cell-phone software and portable vapor-condensation instrument, for obtaining images and analyzing quantitative data..The goal of this project is to develop bio-diagnostic technology with low cost, high sensitivity, selectivity, high-throughput and fast detection. We will explore innovative detection-mechanism of bio-chips based on vapor condensation, as well as develop corresponding portable instruments. This project possesses great value of practical application and broad markets.

重大疾病（如癌症）的早期筛查与诊断是降低疾病死亡率的关键。为满足临床对低成本、高通量、高灵敏度的快速检测技术的迫切需求，本项目创新地将银染技术(<15 min)与汽凝法(<1 min)集成到生物芯片上，以实现快速检测。利用银染生成的微纳米银颗粒改变了检测点的亲水性和颜色，将生物特异性识别转化为汽凝信号输出，形成双级信号放大，可极大地提高灵敏度并节约检测时间。我们将实现对生物标志物如基因突变、miRNA、小分子、离子、蛋白，甚至细胞的快速高通量筛选。检测结果可通过裸眼识别，极大的降低了成本，满足资源匮乏地区需求。为了增加芯片检测的灵活性，满足不同检测环境的需求，我们将开发手机软件以及便携式汽凝仪，采集检测结果并定量分析。.本项目以实现快速、低成本、高通量的分子诊断为目标，开展基于汽凝法的生物芯片检测机制的革新研究以及配套的便携式仪器的研制。该课题具有较强的实际应用价值和广阔的市场前景。

本项目以实现低成本、高灵敏度、高通量和快速的生物分子诊断为目标，开展了新型生物芯片检测技术研究，我们创新地将等温核酸扩增技术、银染技术、目标物引起的DNA连接技术等集成到生物芯片上，将分析物的出现转化为芯片表面的亲疏水属性的改变，以水汽凝结作为检测的报告信号，实现了仅仅通过水汽凝结检测P53以及KRAS等相关基因点突变。为了增加芯片检测的灵活性，满足不同检测环境的需求，开发了便携式的汽凝仪和基于Android手机的定量程序，可以自动识别检测位点以及灰度定量分析，极大的降低了成本，满足资源匮乏地区需求。此外，开发了新型纸质微流控制作方法，降低了开发成本和操作复杂性，实现了低成本、高通量、高精度的小分子的肉眼检测。目前相应方法、设备和软件都申请了相关专利，并正被应用于开发疾病诊断相关生物标志物的检测技术研究中。本项目的实施对于设计新型生物传感器及其在医疗检测领域的应用具有较高的理论和实际应用价值。项目产生的研究成果主要以学术论文形式发表，以第一作者在国内外核心学术期刊上发表SCI、EI及核心论文5篇，申请专利5项，撰写国际著名出版社WILEY-VCH出版的章节1部，参与撰写实验教材1部。多名博士生、硕士生及本科生参与该项目的研究。