State Key Laboratory of Agricultural Microbiology, College of Science, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
State Key Laboratory of Agricultural Microbiology, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, 430070, People's Republic of China.
Mikrochim Acta. 2019 Aug 30;186(9):661. doi: 10.1007/s00604-019-3743-8.
The authors describe an improved lateral flow assay based on (a) the use of catalytic hairpin assembly (CHA), and (b) on signal amplification performed at the interface of gold nanoparticles (AuNPs). The combination of the amplification capability of the CHA reaction and the unique optical properties of AuNPs results in an assay that has a sensitivity that is improved by more than two orders of magnitude. MicroRNA-21 was employed as a model analyte to prove the concept. The presence of microRNA-21 triggers the self-assembly of two hairpin DNAs into double stranded DNA and exposing biotin molecules on the surface of AuNPs. Hence, the target becomes recycled and the signal is strongly amplified. The AuNPs carrying biotin are captured on the test line of the strip to display a red zone. This enables the visual recognition of microRNA without the need for any instrumentation. The fast quantitation of microRNA via the red band intensity is accomplished with the help of software, and the limit of detection is 0.89 pM. The enhanced lateral flow assay was employed to the determination of microRNA-21 in cell extracts and spiked serum samples. Graphical abstract A lateral flow assay for microRNA is described with a detection limit that is improved by two orders of magnitude. It is based on catalytic hairpin assembly (CHA) signal amplification performed at the interface of gold nanoparticles.
作者描述了一种基于 (a) 催化发夹组装 (CHA) 的改进型侧向流动分析,和 (b) 在金纳米粒子 (AuNPs) 界面进行信号放大的侧向流动分析。CHA 反应的放大能力和 AuNPs 的独特光学性质的结合,使得该分析方法的灵敏度提高了两个数量级以上。微小 RNA-21 被用作模型分析物来证明这一概念。微小 RNA-21 的存在触发了两条发夹 DNA 自组装成双链 DNA,并在 AuNPs 表面暴露生物素分子。因此,目标被回收,信号被强烈放大。携带生物素的 AuNPs 被捕获在条带的测试线上,显示出红色区域。这使得无需任何仪器即可进行微小 RNA 的可视化识别。借助软件,可以快速定量通过红色带强度进行微小 RNA 的定量,检测限为 0.89 pM。增强型侧向流动分析用于测定细胞提取物和加标血清样品中的微小 RNA-21。