HCCO纳米反应器集成电化学纸基分析装置用于抗EBNA1 IgA检测以筛查鼻咽癌

文献信息

中文摘要

背景： 抗EB病毒核抗原1免疫球蛋白A（抗EBNA1 IgA）的血清学检测在临床上与鼻咽癌筛查相关，但传统免疫测定依赖集中式仪器和多步操作。方法：在此，我们开发了一种一次性折纸电化学纸基分析装置，集成了中空C@CuO-CeO2纳米反应器（HCCO-ePAD），用于无标记检测抗EBNA1 IgA。HCCO被设计为多功能传感界面，其中中空碳骨架支持电荷和质量传输，含铜结构域提供内在阴极氧化还原响应，表面羧基功能化可实现EBNA1的共价固定。结果：抗EBNA1 IgA与表面结合的EBNA1结合形成导电性差的免疫复合物，限制界面电子转移，导致差分脉冲伏安电流呈浓度依赖性下降。在优化条件下，HCCO-ePAD在0.1至500 ng mL-1范围内呈线性响应，检测限为0.029 ng mL-1，并具有良好的选择性、重复性、重现性和储存稳定性。使用15例鼻咽癌患者和15例健康对照的血清样本进行初步评估，得到受试者工作特征曲线下面积为0.969（95%置信区间，0.918-1.000）。HCCO-ePAD结果进一步与使用商业酶联免疫吸附测定（ELISA）获得的结果进行了比较。结论：这项工作展示了一种内在氧化还原纳米反应器界面，将电化学转导和生物受体固定整合在一次性折纸纸平台上，用于分散式血清学分析。

英文摘要

Serological detection of anti-Epstein-Barr virus nuclear antigen 1 immunoglobulin A (anti-EBNA1 IgA) is clinically relevant to nasopharyngeal carcinoma screening, but conventional immunoassays rely on centralized instrumentation and multistep procedures. Here, we developed a disposable origami electrochemical paper-based analytical device integrating hollow C@CuO-CeO2 nanoreactors (HCCO-ePAD) for label-free anti-EBNA1 IgA detection. HCCO was designed as a multifunctional sensing interface in which the hollow carbon framework supports charge and mass transport, Cu-containing domains provide an intrinsic cathodic redox response, and surface carboxyl functionalization enables covalent EBNA1 immobilization. Binding of anti-EBNA1 IgA to surface-bound EBNA1 formed a poorly conductive immunocomplex that restricted interfacial electron transfer, producing a concentration-dependent decrease in the differential pulse voltammetric current. Under optimized conditions, the HCCO-ePAD showed a linear response from 0.1 to 500 ng mL-1, with a detection limit of 0.029 ng mL-1, together with good selectivity, repeatability, reproducibility, and storage stability. Preliminary evaluation using serum samples from 15 patients with nasopharyngeal carcinoma and 15 healthy controls yielded an area under the receiver operating characteristic curve of 0.969 (95% confidence interval, 0.918-1.000). The HCCO-ePAD results were further compared with those obtained using a commercial enzyme-linked immunosorbent assay (ELISA). This work demonstrates an intrinsic-redox nanoreactor interface that integrates electrochemical transduction and bioreceptor immobilization within a disposable origami paper platform for decentralized serological analysis.