摘要
细胞外囊泡(EVs)因承载多种疾病相关蛋白而被认为是提升诊断效能的重要标志物.本文提出了一种基于DNA逻辑门的EVs表面多蛋白联合分析策略,通过单颗粒电感耦合等离子体质谱(SP-ICPMS)对EVs的精准定量分析,实现肝细胞癌(HCC)的早期诊断.本方法设计了可识别EpCAM(上皮细胞黏附分子)、GPC3(硫酸乙酰肝素蛋白聚糖3)和CD63(分化簇63)3种蛋白的适配体-DNA逻辑回路,利用多标志物共定位的方式对EVs进行特异性识别,进而触发滚环扩增反应,生成特征DNA序列并与纳米金探针杂交,实现SP-ICP-MS信号放大检测.该策略显著提升了肿瘤EVs识别的特异性,有效克服了传统方法灵敏度不足及血清基质干扰等难题,检测限可达950 particles·μL^(−1).应用于50例临床样本检测结果显示,EpCAM/GPC3/CD63三阳性表达的EVs亚群可用于区分HCC与肝硬化患者,诊断敏感度为82.4%,特异性为91.3%,优于常规血清甲胎蛋白检测.本研究所提出的SPICP-MS血清EVs分析策略为HCC的早期诊断提供了新思路,同时也推动了SP-ICP-MS技术在精准医学中的进一步应用.
Extracellular vesicles(EVs),carrying multiple disease-related proteins,have emerged as promising markers to enhance diagnostic performance in early hepatocellular carcinoma(HCC)detection.In this study,we propose a DNA logic gate-based strategy for multiplexed protein biomarker-based detection on the surface of extracellular vesicles(EVs),enabling highly sensitive quantification of HCC-derived EVs using single-particle inductively coupled plasma mass spectrometry(SP-ICP-MS)for precise early diagnosis of HCC.The method employs aptamer-based DNA logic circuits capable of recognizing EpCAM(Epithelial Cell Adhesion Molecule),GPC3(Glypican-3),and CD63(Cluster of Differentiation 63)proteins.Upon co-localization of these markers on EVs,the system triggers a rolling circle amplification(RCA)reaction to generate characteristic DNA sequences,which then hybridize with gold nanoparticle probes,thereby enabling signal amplification and subsequent SP-ICP-MS detection.This approach significantly enhances the specificity of tumor EVs recognition,and effectively overcomes the limitations of conventional methods,such as low sensitivity,and interference from complex serum matrices.The detection limit for EVs reaches down to 950 particles·μL^(−1).The EVs subsets with positive expressions of EpCAM,GPC3 and CD63 were used to distinguish patients with hepatocellular carcinoma from those with liver cirrhosis,the diagnostic sensitivity and specificity of triple-positive EVs were 82.4%and 91.3%,respectively,outperforming conventional serum alpha-fetoprotein(AFP)testing.In conclusion,this study proposes a novel serum EVs analysis strategy,offering a new approach for the early diagnosis of HCC and promoting the further application of SP-ICP-MS technology in precision medicine.
作者
吕政贤
杨剑鑫
刘平果
严晓文
王秋泉
LV Zhengxian;YANG Jianxin;LIU Pingguo;YAN Xiaowen;WANG Qiuquan(Department of Chemistry&the MOE Key Laboratory of Spectrochemical Analysis and Instrumentation,College of Chemistry and Chemical Engineering,Xiamen University,Xiamen,361005,China;Department of Hepatobiliary Surgery,Zhongshan Hospital,Xiamen University,Fujian Provincial Key Laboratory for Chronic Liver Disease and Hepatocellular Carcinoma,Xiamen,361004,China;Zhejiang Key Laboratory of Environment and Health of New Pollutants,School of Environment,Hangzhou Institute for Advanced Study,University of Chinese Academy of Sciences,Hangzhou,310024,China;The Affiliated Hospital of Putian University,Putian,351100,China)
出处
《环境化学》
北大核心
2025年第10期3576-3587,共12页
Environmental Chemistry
基金
国家自然科学基金(22193053,21535007,22074127)
国家重点研究与发展计划项目(2022YFF0710200)资助.
