Department of Chemistry , Colorado State University , Fort Collins , Colorado 80523 , United States.
Department of Microbiology, Immunology, and Pathology , Colorado State University , Fort Collins , Colorado 80523 , United States.
Anal Chem. 2018 Jun 19;90(12):7777-7783. doi: 10.1021/acs.analchem.8b02042. Epub 2018 Jun 1.
Viral pathogens are a serious health threat around the world, particularly in resource limited settings, where current sensing approaches are often insufficient and slow, compounding the spread and burden of these pathogens. Here, we describe a label-free, point-of-care approach toward detection of virus particles, based on a microfluidic paper-based analytical device with integrated microwire Au electrodes. The device is initially characterized through capturing of streptavidin modified nanoparticles by biotin-modified microwires. An order of magnitude improvement in detection limits is achieved through use of a microfluidic device over a classical static paper-based device, due to enhanced mass transport and capturing of particles on the modified electrodes. Electrochemical impedance spectroscopy detection of West Nile virus particles was carried out using antibody functionalized Au microwires, achieving a detection limit of 10.2 particles in 50 μL of cell culture media. No increase in signal is found on addition of an excess of a nonspecific target (Sindbis). This detection motif is significantly cheaper (∼$1 per test) and faster (∼30 min) than current methods, while achieving the desired selectivity and sensitivity. This sensing motif represents a general platform for trace detection of a wide range of biological pathogens.
病毒病原体是全球范围内的严重健康威胁,特别是在资源有限的环境下,当前的传感方法往往不够充分且缓慢,加剧了这些病原体的传播和负担。在这里,我们描述了一种基于带有集成的微丝 Au 电极的微流控纸基分析器件的无标记、即时检测病毒颗粒的方法。该器件最初通过生物素修饰的微丝捕获链霉亲和素修饰的纳米颗粒来进行表征。与经典的静态纸基器件相比,由于增强了质量传输和修饰电极上的颗粒捕获,微流控器件的检测限提高了一个数量级。使用抗体功能化的 Au 微丝对西尼罗河病毒颗粒进行电化学阻抗谱检测,在 50 μL 的细胞培养液中实现了 10.2 个颗粒的检测限。在添加过量非特异性靶标(辛德毕斯)时,未发现信号增加。与当前方法相比,这种检测模式的成本显著降低(∼每测试 1 美元),速度更快(∼30 分钟),同时实现了所需的选择性和灵敏度。这种传感模式代表了广泛生物病原体痕量检测的通用平台。