Department of Health & Environment, Nano Systems, Austrian Institute of Technology GmbH, Donau-City-Street 1, 1220 Vienna, Austria.
Lab Chip. 2009 Dec 21;9(24):3549-56. doi: 10.1039/b914738a. Epub 2009 Oct 12.
Rapid detection of viral contamination remains a pressing issue in various fields related to human health including clinical diagnostics, the monitoring of food-borne pathogens, the detection of biological warfare agents as well as in viral clearance studies for biopharmaceutical products. The majority of currently available assays for virus detection are expensive, time-consuming, and labor-intensive. In the present work we report the creation of a novel micro total analysis system (microTAS) capable of continuously monitoring viral contamination with high sensitivity and selectivity. The specific interaction between shape and surface chemistry between molecular imprinted polymer (MIP) and virus resulted in the elimination of non-specific interaction in the present sensor configuration. The additional integration of the blank (non-imprinted) polymer further allowed for the identification of non-specific adsorption events. The novel combination of microfluidics containing integrated native polymer and MIP with contact-less dielectric microsensors is evaluated using the Tobacco Mosaic Virus (TMV) and the Human Rhinovirus serotype 2 (HRV2). Results show that viral binding and dissociation events can be readily detected using contact-less bioimpedance spectroscopy optimized for specific frequencies. In the present study optimum sensor performance was achieved at 203 kHz within the applied frequency range of 5-500 kHz. Complete removal of the virus from the MIP and device reusability is successfully demonstrated following a 50-fold increase in fluid velocity. Evaluation of the microfluidic biochip revealed that microchip technology is ideally suited to detect a broader range of viral contaminations with high sensitivity by selectively adjusting microfluidic conditions, sensor geometries and choice of MIP polymeric material.
快速检测病毒污染仍然是与人类健康相关的各个领域的一个紧迫问题，包括临床诊断、食源性病原体监测、生物战剂检测以及生物制药产品的病毒清除研究。目前大多数用于病毒检测的方法都昂贵、耗时且劳动强度大。在本工作中，我们报告了一种新型微全分析系统（microTAS）的创建，该系统能够以高灵敏度和选择性连续监测病毒污染。分子印迹聚合物（MIP）与病毒之间的形状和表面化学之间的特异性相互作用导致在本传感器配置中消除了非特异性相互作用。空白（非印迹）聚合物的额外集成进一步允许识别非特异性吸附事件。使用烟草花叶病毒（TMV）和人类鼻病毒 2 型（HRV2）评估了包含集成天然聚合物和 MIP 的微流控和非接触式介电微传感器的新颖组合。结果表明，使用针对特定频率进行优化的非接触式生物阻抗谱可以轻松检测到病毒结合和解离事件。在本研究中，在 5-500 kHz 的应用频率范围内，在 203 kHz 下实现了最佳传感器性能。在流体速度增加 50 倍后，成功地从 MIP 和器件中完全去除了病毒，并实现了器件的可重复使用性。微流控生物芯片的评估表明，微芯片技术通过选择性地调整微流控条件、传感器几何形状和 MIP 聚合物材料的选择，非常适合以高灵敏度检测更广泛范围的病毒污染。
