基于CRISPR-Cas系统的牙龈卟啉单胞菌纸基微流控芯片检测平台

牙周炎不仅影响口腔健康，还与糖尿病、心血管疾病、类风湿性关节炎等疾病之间存在关联。牙龈卟啉单胞菌（Porphyromonas gingivalis）是与慢性牙周炎关系最密切的致病菌。可靠的早期牙周病原检测能为患者的健康带来众多的益处。目前常用的牙龈卟啉单胞菌检测方法都因存在某些缺点而无法普遍应用于牙龈卟啉单胞菌的“椅边”诊断。CRISPR-Cas系统能够特异性的识别和降解靶核酸序列。被开发为高特异性和高灵敏度的核酸检测工具。纸基微流控芯片（paper-based microfluidic devices，μPADs）是以纸为反应场所的微流控可手持设备，天生具有成本低、可折叠、易于操作和便于处理等特点。本项目拟将CRISPR-Cas与μPADs相结合，构建一套从DNA提取到结果输出（荧光/肉眼可视化）一系列流程一体化的检测平台，针对牙龈卟啉单胞菌进行快速、精准的临场检测。

Periodontitis not only affects oral health, but also is associated with diabetes, cardiovascular disease, rheumatoid arthritis and other diseases. Porphyromonas gingivalis is the most closely related pathogen to periodontitis. Reliable early periodontal pathogen detection can not only effectively prevent periodontitis, but also bring more benefits to the health of patients. At present, the commonly used detection methods of Porphyromonas gingivalis can not be widely used in the "chair edge" diagnosis of Porphyromonas gingivalis because of some shortcomings. The CRISPR-Cas system can specifically recognize and degrade target nucleic acid sequences. It has been developed as a nucleic acid detection tool with high specificity and sensitivity. Paper-based microfluidic chip (paper-based microfluidic devices, μPADs) is a microfluidic handheld device with paper as the reaction site, which is inherently low-cost, foldable, easy to operate and easy to handle. This project intends to combine CRISPR-Cas with μPADs to construct a set of detection platform which can carry out a series of detection processes of DNA extraction-target sequence isothermal amplification-target sequence identification-result output (fluorescence / naked eye visualization) for rapid and accurate point-of-care test of Porphyromonas gingivalis.