高灵敏检测肺癌外周血中EGFR基因突变的数字PCR芯片研究

项目名称： 高灵敏检测肺癌外周血中EGFR基因突变的数字PCR芯片研究

项目编号： No.61271162

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 无线电电子学、电信技术

项目作者： 赵建龙

作者单位： 中国科学院上海微系统与信息技术研究所

项目金额： 86万元

中文摘要： 肺癌是世界上发病率和死亡率最高的恶性肿瘤。表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKIs）作为一种分子靶向药物，可以提高晚期非小细胞肺癌患者的生存率和生活质量。但这类药不是对所有肺癌患者都有效，EGFR的基因突变与其药效及耐药性密切相关，准确、高效地对患者尤其是晚期患者外周血进行EGFR基因突变测定，对患者进行有针对性的靶向药物治疗具有重要的临床意义。鉴于外周血中肿瘤细胞产生的循环DNA量比较少、突变DNA分子更少的特点，通过制作含有纳升级乳滴微反应器的数字PCR芯片，将待测DNA分子分散到上万个纳升级的乳滴中，结合位点特异性扩增及Taqman荧光探针，通过检测每一乳滴的信号获得EGFR突变情况，利用建立的EGFR基因突变检测方法，研究外周血与组织标本EGFR基因突变的一致性，在此基础上探索一种新的外周血EGFR基因突变的高灵敏、高特异检测方法，为肺癌靶向药物的治疗提供依据。

中文关键词： 液滴式数字PCR；微流控芯片；循环肿瘤DNA；EGFR基因检测；肺癌

英文摘要： Lung cancer is one of the most malignant tumors with high morbidity and mortality in the world. Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs), as a molecular targeted drug, improve the survival and quality of life for advanced non-small cell lung cancer patients. As the best biomarker, EGFR gene mutation is correlated with its efficacy and drug resistance. So, whether the mutation of EGFR gene is exist in lung cancer patients should be detected firstly. The TKIs can be as the first line treatment in these patients at advanced stages with EGFR mutation. Most of patients at the advanced stages had lost chances of surgery, the tissues samples of these kinds of patients can't get. The peripheral blood is easy to get for these kinds patients. However, the mount of circulating DNA in peripheral blood is relatively low and the mutant DNA is less, which is difficult to detect. The digital PCR chip was conduct to detect the few mutants. The DNA molecules will be divided into thousands of emusion droplets with nanoliter, in which there is zero, one,or two copy of DNA. These DNA molecules were amplified with allele specific primer amplification and detected with TaqMan probes with fluorescent. The fluorescent signal of every emusion droplets will be detected or scanned with fluorescent microscopy

英文关键词： droplet digital PCR；microfluidic chip；ctDNA；EGFR detection；lung cancer