用于外周血循环肿瘤细胞高灵敏检测石墨烯微芯片传感器的研究

Circulating tumour cells(CTC) in the peripheral blood are derived from the tumor tissue, which play a significant role in tumor detection, tailored therapy, treatment evaluation, prognosis monitoring, etc. Owing to the extremely low number of CTCs presenting in peripheral blood, it’s technically challenged to sensitively and rapidly measure defined CTCs in peripheral blood. Now the method based on optical immunofluorescent staining detection is mainly utilized to detect CTC, however, this approach has complex operations, exorbitant price and it’s difficult to apply in the clinical. We intend to develope a graphene microchipsensor for the detection of CTC in peripheral blood based on the unique advantage of the excellent electric conductivity of graphene’s in biochemical detection, in which CTCs are captured by functionalized graphene, and further quantitatively detected according to the resistance change on graphene surface. A helical inductive coil is designed to couple with a reader unit by a coil antenna in the reader system and the changes of frequency characteristic in the graphene-based resonant network could be wireless monitored by the reader unit for the further quantitative analysis of CTC. This method could improve the detection sensitivity, resolution and resistance to interference significantly. This project will reveal the sensitivity and accuracy of CTC detection by the establishment of the graphene-based microchip sensor. Moreover, we will study the clinical significance of CTCs on the evaluation and prediction to the treatment responses in lung cancer and colorectal cancer. This study will provide an efficient, convenient methodology for further investigation of early response and prognosis evaluation in an efficient, convenient way，which have important values and significance in clinical application.

外周血中的循环肿瘤细胞（circulating tumour cell, CTC)来源于肿瘤组织，对肿瘤的检测、疗效评估及预后监测等具有重要意义。由于CTC在血液中含量非常少,如何高灵敏、快速及准确地测定外周血中CTC含量，面临技术上的挑战。目前CTC检测主要利用免疫荧光染色的光学检测手段，操作繁琐，价格昂贵，难以在临床上推广。本项目提出基于石墨烯电学特性检测CTC的微芯片传感系统，根据石墨烯表面捕获细胞后电阻的变化，采用电容-电感对构建射频共振网络，通过读出电路测量传感器谐振网络频率特性变化实现对CTC定量检测，可显著提高检测灵敏度、分辨率和抗干扰能力。本项目将解决微量CTC高灵敏及准确检测等关键问题基础上，建立CTC电学定量检测方法，构建石墨烯微芯片传感器，并用于临床样本的分析验证。本项目的研究为建立一种高效、方便的肿瘤检测及预后监测的新模式提供新思路，具有重要的临床应用价值和意义。

癌症是我国死亡率最高的疾病之一，癌症的发生率和死亡率目前呈逐年上升趋势，而 90 %的死亡源于肿瘤的转移和复发。研究表明，在肿瘤转移的早期或未发生转移时，体内一些肿瘤标记物就已发生变化，比如蛋白肿瘤标志物以及循环肿瘤细胞（circulating tumor cells, CTCs）等。液体活检技术是基于微创体液检测方法，具有显着改变癌症患者治疗策略的巨大潜力，为患者个体提供极其强大且可靠的非侵入式的实时临床诊断手段。近年随着个性化医疗以及精准医学的发展，液体活检技术被广泛地用于肿瘤的早期预警、筛查以及预后的监测。.本项目的研究聚焦了液态活检中肿瘤蛋白标志物和CTC两类标志物，并且结合了二维纳米材料、微流控以及无线射频技术等多个学科技术，实现了多种肿瘤标志物的检测与分析，提供了基于微纳器件结合微流控技术的液态活检新方法。本项目的主要研究内容：（1）石墨烯薄膜进行自组装修饰以及石墨烯电学生物传感器的构建；（2）构建分离、捕获CTC 以及单个CTC分析的微芯片平台；（3）设计高灵敏度的传感器电容和作为天线耦合电感的RCL谐振网络无线射频芯片，通过信号读取线圈，可直接利用RCL谐振网络无线射频芯片实现CTC的检测。.本项目在石墨烯表面修饰以及石墨烯生物传感器的构建方面，针对石墨烯表面修饰的问题，提出了新型的石墨烯表面修饰以及石墨烯场效应管器件的构建方法；设计了可用于单个CTC细胞分选、捕获以及原位分析的微流控芯片；通过结合单个CTC细胞分选芯片和无线射频芯片实现了CTC的检测，这种基于细胞介电特性检测的方法在未来可用于不同肿瘤细胞的分选，为肿瘤细胞的分选或者分型提供一种新的技术。