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性病原体暴露至关重要，无论是来自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志物，可用于识别最近或过去的感染因子暴露，抗体分析方法对于含有可检测的病毒或病原体遗传物质的可忽略量的样品尤其重要。临床中常用的抗体反应测量方法通常受到关注单一病原体、几个耗时的执行步骤、数据采集的周转期较慢、特异性和敏感性问题以及结果可变等问题的阻碍。我们建议开发一种简单的微流体装置，它将整合抗体诊断所需的许多复杂任务，从处理手指刺血样本到识别特定感染因子，利用微流体和纳米流体的独特物理特性来提高检测性能。我们的研究将侧重于对公共卫生具有最大潜在影响的两类病原体引起的感染的即时诊断：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）。我们设想微流控装置的微阵列格式可以迅速扩展到包括大量其他生物制剂的测试。该合作团队将由来自传染病研究所生物传感器和蛋白质微阵列实验室以及马里兰大学微机电系统和微流体实验室的研究人员组成。项目团队在传染病研究、MEMS、微流体系统和基于微阵列的技术方面拥有独特的专业知识，用于高通量检测人类抗体与病原体特征蛋白的相互作用。临床和兽医感染病例的血清将用于对该设备进行广泛评估。我们的方法将只需要纳升数量的血清进行分析，并将导致开发一种低成本、一次性设备，用于快速、灵敏地检测症状前、症状或恢复期传染病的生物标志物。

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.