Integrated polymer microfluidic device for multiplex diagnosis of viral infection

描述（由申请人提供）：快速准确的感染诊断对于管理高毒性病原体的潜在暴露至关重要，无论是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暴露于传染性病原体，抗体分析方法对于含有可忽略量的可检测病毒或病原体遗传物质的样品尤其重要。在临床中用于测量抗体应答的常用方法通常受到以下因素的阻碍：关注单一病原体、若干耗时的执行步骤、用于数据采集的缓慢周转时间、特异性和灵敏度问题以及可变结果。我们建议开发一种简单的微流控设备，该设备将集成基于抗体的诊断所需的许多复杂任务，从处理手指针刺血液样本到识别特定传染病原的暴露，利用微流控和纳米流控的独特物理特性来增强检测性能。我们的研究将侧重于对公共卫生影响最大的两种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。该合作小组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州大学微机电系统（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，微流体系统和基于微阵列的技术的独特专业知识，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器械的广泛评价。我们的方法将只需要纳升量的血清进行分析，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，症状或恢复期生物标志物。 公共卫生相关性：这项拟议中的研究将产生一种简单、低成本的设备，用于快速诊断由许多对公众健康很重要的病毒引起的感染。这种小型一次性塑料装置将针对在实验室方法方面经验很少的从业者。

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. PUBLIC HEALTH RELEVANCE: The proposed research will result in a simple, low cost device for the rapid diagnosis of infections caused by many viruses that are important to public health. This small, disposable plastic device will be targeted for practitioners with very little experience in laboratory methods.