分子靶向治疗是目前大肠癌最有效的治疗策略，但如果病人的某些基因存在突变，则靶向治疗无效，因此，亟需发展高效的检测方法筛查病人的基因突变状态，以准确指导临床个体化的用药。目前采用的突变检测方法存在灵敏度低、样品用量大、操作繁琐费时等问题，而且难以对相关的多个基因进行同时检测。为此，前期工作将基因突变检测方法PCR-RFLP中的电泳分离操作移植到微流控芯片上进行，显著提高了突变检测灵敏度。为了进一步提高其对微量样品的快速检测能力和多指标平行分析能力，本研究拟利用微流控液滴技术能够连续操控多个不同微反应单元的优势和高通量分析能力，创建基于液滴的集成化PCR-RFLP系统，即自动化完成基于液滴的微量DNA样品前处理；连续完成多样品阵列式液滴PCR扩增和酶切反应；进行液滴内多样品的连续自动进样和毛细管电泳分析。应用于大肠癌肿瘤组织和血液中靶向治疗相关基因的检测，为指导肿瘤个体化治疗提供高效技术平台。

Molecular targeted therapy is currently the most effective way of treating metastatic colorectal cancer. Because the patients harboring mutations in genes such as K-ras cannot benefit from the treatment, it is necessary to develop highly effective detection methods to screen the gene mutation status of the patients to facilitate personalized treatment. However, methods currently in use are not able to reveal mutations efficiently due to low sensitivity, high sample and time consumption, and failure to test therapy-related genes simultaneously. To this end, in previous work, we transplanted PCR-RFLP, a method for detecting gene point mutations, onto microfluidic chip and achieved a significant increase in sensitivity. To further enhance the ability to rapidly detect and parallel analyze trace amount of samples, this study focuses on developing an integrated PCR-RFLP platform based on microfluidics droplet by combining whole analysis process together, including DNA extraction system which can perform mini DNA preparation automatically, multiple droplet array which can sequentially fulfill PCR amplification and enzyme digestion processes for different samples, and capillary electrophoresis system which can continuously introduce and analyze samples in droplets. The established platform can be utilized to test mutations of several genes related to targeted therapy in tumor tissue and paired serum of colorectal cancer patients, providing additional insights to the personalized therapy of cancer.