Department of Clinical Laboratory, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, Hangzhou, 310014, China.
Key Laboratory of Clinical Laboratory Diagnostics (Ministry of Education), College of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016, China.
Biosens Bioelectron. 2019 Oct 1;142:111548. doi: 10.1016/j.bios.2019.111548. Epub 2019 Jul 30.
Antimyeloperoxidase (anti-MPO) is regarded as one of the most important circulating autoantibodies for anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitides (AAVs). Hence, it is crucial for highly sensitive detection of anti-MPO to monitor efficacy of AAVs in clinical diagnosis. Herein, a highly sensitive electrochemiluminescence (ECL) immunosensor for anti-MPO detection was constructed by combining reduced graphene oxide-supported PtCo nanocubes hybrids (PtCo@rGO) with hybridization chain reaction (HCR) as signal amplification. Multiple ECL luminophores (Dox-ABEI) prepared by cross-linking of N-(aminobutyl)-N-(ethylisoluminol) (ABEI) and doxorubicin (Dox) were intercalated into dsDNA products of HCR, achieving the effective immobilization of ECL luminophores to obtain strong ECL emission. Benefiting from the efficient catalytic activity of PtCo@rGO toward HO, the massive the superoxide radical (O-) were generated to further react with ABEI for ECL emission. Thus, the designed ECL immunoassay for anti-MPO detection exhibited excellent sensitivity of a concentration variation from 50 fg/mL to 1 ng/mL and a detection limit of 15.68 fg/mL. Importantly, this work proposed an enzyme-free ECL immunoassay with high sensitivity, excellent specificity for protein detection in clinical diagnosis.
抗髓过氧化物酶（anti-MPO）被认为是抗中性粒细胞胞质抗体（ANCA）相关性血管炎（AAV）中最重要的循环自身抗体之一。因此，高灵敏度检测 anti-MPO 对于临床诊断中监测 AAV 的疗效至关重要。在此，通过将负载 PtCo 纳米立方体的还原氧化石墨烯（PtCo@rGO）与杂交链式反应（HCR）结合，构建了一种用于抗-MPO 检测的高灵敏度电化学发光（ECL）免疫传感器。通过交联 N-(氨丁基)-N-(乙基异鲁米诺)（ABEI）和阿霉素（Dox）制备的多个 ECL 发光体（Dox-ABEI）被插入 HCR 的 dsDNA 产物中，实现了 ECL 发光体的有效固定，从而获得了强的 ECL 发射。得益于 PtCo@rGO 对 HO 的高效催化活性，大量超氧自由基（O-）生成，进一步与 ABEI 反应进行 ECL 发射。因此，所设计的用于抗-MPO 检测的 ECL 免疫分析表现出优异的灵敏度，浓度变化范围为 50 fg/mL 至 1 ng/mL，检测限为 15.68 fg/mL。重要的是，这项工作提出了一种无酶 ECL 免疫分析方法，具有高灵敏度、优异的特异性，可用于临床诊断中的蛋白质检测。
