基于微流控芯片的循环肿瘤细胞无标记分选及基因检测新方法研究

本项目拟设计构建多级模块化微流控芯片系统，用于外周血中循环肿瘤细胞（CTC）的快速无标记分离，及高灵敏基因检测。癌症是严重威胁人类生命健康的重大疾病之一，肿瘤细胞脱落、侵袭并进入血液循环，到达全身组织、影响器官功能是造成癌症相关死亡的主要原因。特异、敏感地检测血液中极微量的CTC有助于肿瘤早期诊断和复发转移监控，并为肿瘤治疗提供新的靶点和策略。芯片集成（1）无标记肿瘤细胞分离富集模块；（2）芯片在线核酸提取模块；及（3）核酸等温扩增和读出模块，配套小型便携式的仪器，实现芯片的自动操作及结果判读。多级模块化微流控芯片系统具有结构简单，高通量、无复杂仪器要求、操作简便等优点，有利于形成一套完整的基于CTC的癌症早期诊断及疗效评价的整套实验方案和标准，其不仅具有重要的科学意义，而且将有效推动我国微流控芯片技术抢占国际科技竞争的制高点。

This proposal aims at developing a multi-stage microfluidic system for a rapid and continuous separation of circulating tumor cells (CTCs) from peripheral blood in a label-free manner, and on-chip pretreatment, amplification, and detection of nucleic acids in isolated CTCs. CTCs shed from either primary or metastatic cancers have been identified in the peripheral blood of patients, and are often associated with cancer metastasis and tumor recurrence. The majority of deaths from cancer in human patients are caused by metastasis, suggesting a need to precisely isolate and detect CTCs from whole blood. The proposed microfluidic system integrates: (1) a label-free CTCs isolation module; (2) a nucleic acid pretreatment module; and (3) an on-chip Loop-mediated isothermal amplification (LAMP) module. The operation of the microfluidic chip and the readout of results are automated by designing a custom-made portable instrument. As the proposed microfluidic system carries many advantages such as disposability, label-free, high-throughput, easy operation, low cost, and reduced cross contamination and biohazard risks, we expect this system to have an impact on fundamental and clinical studies of circulating tumor cells.