基于纳米滚环扩增协同表面增强拉曼光谱法检测lncRNA用于急性肾损伤精准诊断

长链非编码核糖核酸（lncRNA）作为疾病的关键调控因子，在急性肾损伤生理病理过程中起重要作用。本项目拟结合表面增强拉曼光谱与超支化滚环扩增技术，合成多种结构的高灵敏拉曼探针，同时针对不同靶序列设计特异性挂锁探针，借助一体化微流控分析平台，在芯片内实现超支化滚环扩增与拉曼检测。通过扩增产物促进SERS探针形成纳米团簇，协同拉曼信号进行放大，从而实现低扩增数下的有效检测。构建预临床肾损伤模型用于相关lncRNA的检测，确定最优的探针与芯片设计，在组织、体液多层面上跟踪lncRNA变化，验证相关lncRNA在急性肾损伤精准诊断中的作用，同时深入探究该检测手段的临床适用性。在检测时间、可重复性、稳定性与灵敏度上实现突破，提高实际检测中急性肾损伤诊断结果的准确性。研究结果预期为临床急性肾损伤的检测提供快速高灵敏的液体活检新方法，进而为急性肾损伤的早期精准诊断、术中以及预后的监测提供数据支撑。

Considerable interest has been aroused in long non-coding RNA (lncRNA) due to its vital roles in various pathophysiological processes of acute kidney injury (AKI). This project intends to combine surface-enhanced Raman spectroscopy and hyper-branched rolling circle amplification, to synthesize highly sensitive Raman probes of various structures, and to design specific padlock probes for the detention of different target sequences. Highly sensitive detection of target lncRNA will be performed by hyperbranched rolling circle amplification and Raman detection in the integrated microfluidic analysis platform. The amplification product promotes the formation of nanoclusters. Thus amplifying the Raman signal, and achieving effective detection at low amplification numbers. A pre-clinical kidney injury model will be designed for the detection of the AKI related lncRNA. By optimizing SERS probes, chip design and tracking related lncRNA changes on tissues and body fluids, This will help for further understanding related lncRNA in the accurate diagnosis of acute kidney injury and stretch out the in-depth clinical capability of the proposed method. A breakthrough will be achieved in detection time, repeatability, stability, and sensitivity to improve the accuracy of the AKI diagnosis in practical detection. The results are expected to establish a new method of rapid and highly sensitive lncRNA humoral biopsy in complex environments, hoping to provide a research basis for early accurate diagnosis of clinical acute kidney injury, timely feedback of intraoperative data and effective postoperative monitoring.