呼出气体冷凝液中痕量生物标志物的表面增强电化学发光成像分析

Exhaled breath condensates (EBCs) contain a multiple of biomarkers, such as carcinoembryonic antigen, tumor necrosis factor, endothelin-1, interleukin, and so on. These biomarkers are vital for the diagnosis of corresponding diseases. However, the concentrations of these biomarkers are normally 2000 to 10000 times lower than those in human body fluid. Thus those commercially available apparatus currently existed in the hospital usually cannot detect these biomarkers in EBCs. This has become the bottle-neck of using EBCs for clinical diagnosis of diverse diseases. In our project, we are going to combine the microfluidic technique, the immunoassay technique, and the surface enhanced electrochemiluminescence imaging (SEECL-I) technique to develop an ultrasensitive and high throughput analytical method for the detection of biomarkers in EBCs. This project are intended to realize the following three goals: 1) To solve the insufficient sensitivity problem of current commercially available approaches; 2) To screen new biomarkers in EBCs; and 3) To establish a simple, high efficient, noninvasive and accurate method for the diagnosis of disease in respiratory system. Meanwhile, our method would also provide a new analytical method for other researchers in relevant fields, and we believe this project would push the application of EBCs for clinical diagnosis of different kind of respiratory diseases.

呼出气体冷凝液（EBCs）中含有癌胚抗原、肿瘤坏死因子、内皮素-1、白介素等一系列生物标志物，对呼吸系统疾病的诊断具有非常重要的参考价值。然而，由于这些标志物在EBCs中的浓度要比人体液中低2000-10000倍，因此目前医院中大部分检测手段无法从EBCs中检出相应标志物，这已经成为了使用EBCs进行临床诊断的重要瓶颈之一。本项目拟将微流控芯片技术、免疫分析技术与表面增强电化学发光成像技术（SEECL-I）相结合，开发超高灵敏度、高选择性及高通量的EBCs分析新方法。本项目拟实现以下三个目标：1) 解决现有EBCs分析方法中灵敏度不够的问题； 2) 在EBCs中筛选出新的特异性好、敏感度高的生物标志物；3) 建立简单、高效、无创、精准的呼吸系统疾病诊断新方法。同时，本项目的成功实施也将为相关领域的研究人员提供一种强有力的分析手段，从而为推动EBCs进入常规临床诊断做出贡献。

本项目将免疫分析技术与表面增强电化学发光成像技术（SEECL-I）相结合，开发系列超高灵敏度、高选择性及高通量的EBCs分析新方法。主要完成了以下几项研究任务：.1).研制了一套电致化学发光暗场成像分析系统，该系统由一台暗视野显微镜、一台光谱仪、一部EMCCD相机及一部彩色CCD相机组成，能用于单纳米粒子水平的电致化学发光成像分析及光谱测量；.2).利用我们自研的ECL成像分析系统，我们在呼出气体冷凝液中成功鉴定出五种疾病标志物，分别为癌胚抗原（CEA）、神经元特异性烯醇化酶（NSE）、鳞状上皮细胞癌抗原（SCC）、miRNA-21和miRNA-155；.3）建立了一系列在呼出气体冷凝液中分析上述疾病标志物的分析方法。.项目研究成果在本领域权威期刊上发表研究论文26篇，其中SCI影响因子大于5的论文22篇，以本项目基金号为第一基金标注的论文20篇；授权中国发明专利5项，实用新型专利1项。