Li Tao, Ou Gaozhi, Chen Xuliang, Li Zheyu, Hu Rui, Li Ying, Yang Yunhuang, Liu Maili
State Key Laboratory of Magnetic Resonance and Atomic Molecular Physics, Wuhan National Laboratory for Optoelectronics, National Centre for Magnetic Resonance in Wuhan, Wuhan Institute of Physics and Mathematics, Innovation Academy for Precision Measurement Science and Technology, Chinese Academy of Sciences, Wuhan, 430071, China; University of Chinese Academy of Sciences, Beijing, 10049, China.
School of Sports, China University of Geosciences, Wuhan, 430074, China.
Anal Chim Acta. 2020 Sep 15;1130:20-28. doi: 10.1016/j.aca.2020.07.031. Epub 2020 Jul 28.
Fast and sensitive detection of E.coli O157: H7 is significantly essential for clinical management as well as for transmission prevention during disease outbreaks. Though many types of detection strategies have been implemented for measuring E.coli O157: H7, most of them still rely on complex instruments or tedious/laborious setups, which restrict their applications in resource-limited scenarios. Herein, we introduce an eye-based microfluidic aptasensor (EA-Sensor) for fast detection of E.coli O157: H7 without the assist of any instruments. We demonstrate the perfect coupling of aptamer sensing, hybridization chain reaction (HCR)-amplification and a distance-based visualized readout to quantitatively determine the pathogen concentration. We first used gel-electrophoresis assay to evaluate the system and the results proved that E.coli O157: H7 was well recognized by the aptamer and HCR could increase the signal by about 100 folds. In addition, the Aptamer specificity and signal-amplification ability were verified on the EA-Sensor for sensing E.coli O157: H7 by naked eyes. Furthermore, we demonstrated that E.coli O157: H7 in milk could be accurately and conveniently measured with good performance. With the benefits of operation integration and strategy integration, our EA-Sensor shows advantages of high specificity, easy operation, efficient amplification and visualized readout, which offers a favorable point-of-care tool for E.coli O157: H7 or other pathogen detection in resource-constrained settings.
快速灵敏地检测大肠杆菌O157:H7对于临床管理以及疾病暴发期间的传播预防至关重要。尽管已经实施了多种检测策略来检测大肠杆菌O157:H7，但其中大多数仍依赖复杂的仪器或繁琐的设置，这限制了它们在资源有限场景中的应用。在此，我们介绍一种基于眼睛的微流控适体传感器（EA-Sensor），用于在无需任何仪器辅助的情况下快速检测大肠杆菌O157:H7。我们展示了适体传感、杂交链式反应（HCR）扩增和基于距离的可视化读数的完美结合，以定量确定病原体浓度。我们首先使用凝胶电泳分析来评估该系统，结果证明适体能够很好地识别大肠杆菌O157:H7，并且HCR可以将信号增强约100倍。此外，通过肉眼在EA-Sensor上验证了适体的特异性和信号放大能力，用于检测大肠杆菌O157:H7。此外，我们证明了牛奶中的大肠杆菌O157:H7能够以良好的性能被准确且方便地测量。凭借操作集成和策略集成的优势，我们的EA-Sensor具有高特异性、操作简便、高效扩增和可视化读数的优点，为资源受限环境中大肠杆菌O157:H7或其他病原体的检测提供了一种有利的即时检测工具。
