Agn纳米簇标记CE-ECL方法靶向检测胃癌血浆lncRNAs的应用研究

血浆lncRNAs联合检测对胃癌诊断有重要意义。目前临床上对多种lncRNAs同时检测，存在灵敏度低、假阳性率高的问题。研究证实，毛细管电泳（CE）与电致发光（ECL）联用适于痕量多组分标志物的快速同时检测。项目组前期发现，纳米银兼具增强电泳分离和放大ECL信号双重特性，显著提高检测灵敏度。预实验提示特异性区段的ECL靶向检测能实现lncRNA的特异分析。本项目在此基础上，拟构建毛细管电泳-电致发光（CE-ECL）分析方法，用于血浆中痕量多组分lncRNAs超敏特异检测。采用Agn纳米簇标记响应探针结合lncRNA，经“CE分离+ECL靶向捕获特异性区段”检测，以CE迁移时间-ECL强度二维参数对多种lncRNA同时定量，结合纳米标记、微阵列修饰等技术，从特异性和灵敏度两方面提高多组分同时检测效率，为胃癌早期分子诊断方法及标准的建立提供依据。

The multiple lncRNAs joint assay is significant for the diagnosis of gastric cancer. The present method has a risk of low sensitivity and high false positive. The capillary electrophoresis (CE) coupling electrochemiluminescence (ECL) has been confirmed to fit for the fast and simultaneous analysis of trace multi-lncRNAs. Our previous studies have been suggested that nano-Ag possess dual peculiarity of strengthening the electrophoretic separation and amplifying the ECL signal. And the preliminary experiment suggested the ECL targeted detection for specific sequence segment can profile the lncRNA. Therefore, in this proposal, we will construct the CE-ECL analysis method for the trace multi-lncRNAs in plasma. At first, nano-Ag clusters were utilized to mark the signal probe and conjugated with the lncRNA. Then the multi-lncRNAs conjugates separated in the capillary electrophoresis, followed by ECL targeted detection for specific sequence segment. The multi-lncRNAs were quantified according to the two-dimension parameters of CE migration time versus ECL intensity. Combined with the technologies of nanoparticles label and microarray decoration, the proposed object aimed at improving efficiency of multi-lncRNAs simultaneous analysis from the sensitivity and specificity. This research will provide the molecular diagnosis method for early diagnosis of gastric cancer and offer the lcnRNAs-based diagnostic criteria.