Integrated polymer microfluidic device for multiplex diagnosis of viral infection

描述(由申请人提供)：快速和准确的感染诊断对于管理潜在的高毒力病原体暴露至关重要，无论是发生在生物恐怖主义行为还是自然事件。抗体是非常敏感的血清生物标记物，可用于识别最近或过去接触感染剂的情况，而抗体分析方法对于含有微不足道的可检测到的病毒或病原体遗传物质的样本尤其重要。临床上常用的检测抗体反应的方法通常受制于对单一病原体的关注、几个耗时的执行步骤、数据采集的缓慢周转时间、特异性和敏感性问题以及可变的结果。我们建议开发一种简单的微流控设备，它将集成基于抗体的诊断所需的许多复杂任务，从处理手指刺血样本到识别接触特定感染剂的情况，利用微流体和纳米流体的独特物理性质来提高检测性能。我们的研究将集中于对两种对公共卫生具有最大潜在影响的病原体组造成的感染的护理诊断：虫媒病毒(日本脑炎、基孔肯雅热、登革热、黄热病等)和丝状病毒(埃博拉、马尔堡)。我们设想，微流控设备的微阵列形式可以迅速扩展，以包括对大量其他生物制剂的测试。合作小组将由来自传染病研究所的生物传感器和蛋白质微阵列实验室以及马里兰大学的微电子机械系统(MEMS)和微流体实验室的研究人员组成。该项目团队包括传染病研究、MEMS、微流控系统和基于微阵列的高通量检测人类抗体与病原体标志性蛋白相互作用的技术方面的独特专业知识。临床和兽医感染病例的血清将用于对该设备进行广泛的评估。我们的方法将只需要纳升的血清进行分析，并将导致开发一种低成本的一次性设备，用于快速和灵敏地检测传染病的症状前、症状或恢复期的生物标志物。

DESCRIPTION (provided by applicant): The rapid and accurate diagnosis of infection is critical for managing potential exposures to highly virulent pathogens, whether occurring from an act of bioterrorism or a natural event. Antibodies are very sensitive serum biomarkers that can be exploited to identify recent or past exposures to infectious agents, and antibody analysis methods are especially important for samples containing negligible amounts of detectable virus or genetic material from the pathogen. Commonly practiced methods for measuring antibody responses in the clinic are generally encumbered by a focus on single pathogens, several time-consuming performance steps, slow turn-around time for data acquisition, issues of specificity and sensitivity, and variable results. We propose to develop a simple microfluidic device that will integrate many complex tasks required for antibody- based diagnosis, from processing of finger-prick blood samples to identifying exposures to specific infectious agents, taking advantage of the unique physical properties of micro- and nanofluidics to enhance assay performance. Our studies will focus on point-of-care diagnosis of infections caused by two pathogen groups with the greatest potential impact on public health: arboviruses (Japanese Encephalitis, Chikungunya, Dengue, Yellow Fever and others) and filoviruses (Ebola, Marburg). We envision that the microarray format of the microfluidic device can be rapidly expanded to include tests for an extensive number of other biological agents. The collaborative team will consist of investigators from the Biosensor and Protein Microarray Laboratory from the Research Institute of Infectious Diseases, and the MicroElectroMechanical Systems (MEMS) and Microfluidics Laboratory of the University of Maryland. The project team includes unique expertise in infectious disease research, MEMS, microfluidic systems, and microarray-based technologies for the high-throughput detection of human antibody interactions with signature proteins of pathogens. Sera from clinical and veterinary cases of infection will be used for extensive evaluation of the device. Our approach will require only nanoliter quantities of serum for analysis and will lead to the development of a low cost, disposable device for rapid and sensitive detection of pre-symptomatic, symptomatic or convalescent biomarkers of infectious diseases.