Department of Laboratory Medicine, Peking University Shenzhen Hospital, Shenzhen, 518055, China; Shenzhen Key Laboratory of Micro/Nano Biosensing, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China; Department of Medical Laboratory, Weifang Medical University, Weifang 261053, China.
Shenzhen Key Laboratory of Micro/Nano Biosensing, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China.
Biosens Bioelectron. 2023 Nov 1;239:115623. doi: 10.1016/j.bios.2023.115623. Epub 2023 Aug 19.
Development of specific signal reporters with signal amplification effect are highly needed for sensitive and accurate detection of pathogen. Herein, we design a colorimetric immunosensing nanosystem based on liposome encapsulated quantum dots-sized MnO nanozyme (MnOQDs@Lip) as a signal reporter for ultrasensitive and fast detection of SARS-CoV-2 antigen. The pathogenic antigens captured and separated by antibody-conjugated magnetic beads (MBs) are further connected with antibody-modified MnOQDs@Lip to form a sandwich-like immunocomplex structure. After triggered release, MnO QDs efficiently catalyze colorless 3,3',5,5'-tetramethylbenzidine (TMB) to blue oxidized TMB, which can be qualitatively observed by naked eyes and quantitatively analyzed by UV-Vis spectra or smartphone platforms. By taking advantages of immuno-magnetic separation, excellent peroxidase-like catalytic activity of MnO QDs, and high encapsulation efficiency of MnOQDs@Lip, ultrasensitive detection of SARS-CoV-2 antigen ranging from 0.1 pg/mL to 100 ng/mL is achieved within 20 min. The limit of detection (LOD) is calculated to be 65 fg/mL in PBS buffer. Furthermore, real clinical samples of SARS-CoV-2 antigens can be effectively identified by this immunosensing nanosystem with excellent accuracy. This proposed detection nanosystem provides a strategy for simple, rapid and ultrasensitive detection of pathogens and may shed light on the development of new POCT detection platforms for early diagnosis of pathogens and surveillance in public health.
为了实现对病原体的灵敏、准确检测,高度需要开发具有信号放大效应的特定信号报告分子。在此,我们设计了一种基于脂质体包封的量子点大小 MnO 纳米酶(MnOQDs@Lip)的比色免疫传感纳米系统,作为信号报告分子,用于超灵敏和快速检测 SARS-CoV-2 抗原。通过抗体偶联磁珠(MBs)捕获和分离的病原体抗原,进一步与抗体修饰的 MnOQDs@Lip 连接,形成三明治样免疫复合物结构。触发释放后,MnO QDs 能高效催化无色 3,3',5,5'-四甲基联苯胺(TMB)生成蓝色氧化 TMB,可通过肉眼定性观察和紫外-可见光谱或智能手机平台进行定量分析。利用免疫磁分离、MnO QDs 优异的过氧化物酶样催化活性和 MnOQDs@Lip 的高包封效率,该免疫传感纳米系统可在 20 分钟内实现 SARS-CoV-2 抗原从 0.1 pg/mL 至 100 ng/mL 的超灵敏检测。在 PBS 缓冲液中,LOD 计算值为 65 fg/mL。此外,该免疫传感纳米系统还可有效识别 SARS-CoV-2 抗原的真实临床样本,具有出色的准确性。该检测纳米系统为简单、快速和超灵敏检测病原体提供了一种策略,可能为开发用于病原体早期诊断和公共卫生监测的新型 POCT 检测平台提供思路。