MMDx: A rapid multiplexed matrix code diagnostic for real time epidemiology

7.项目总结/摘要 该建议描述了一种快速多路复用矩阵码诊断的设计、构造和测试， 选择生物防御和新兴病原体。生物攻击的威胁突出表明， 快速护理点诊断，识别感染者体内的特定病原体。感染或有风险 个体可能远离重症监护设施;因此，诊断测试必须易于运输， 储存稳定，使用简单，易于解释。侧流试验（也称为免疫层析 测试或条测试）是一个完善的诊断平台，因为它们的简单性，快速读出，和低 抗体或抗原的定性/半定量检测的成本。然而，缺乏可用点- 传染病的护理诊断代表了生物防御准备的脆弱性， 满足公共卫生需求的差距。我们提出了新的诊断设备设计，包括快速响应 (QR)二维矩阵编码读出，沿着注意化学偶联方法， 提高灵敏度并拓宽侧流装置的多路复用能力。有效的公共卫生 防治工作还需要根据诊断数据了解疾病分布和移动情况。因此，我们认为， 我们提出的多路矩阵诊断（MMDx）设备将是机器可读的，代表一个领域 使用移动的电话技术的具有真实的实时流行病学的集成能力的诊断， 地理定位。我们组建了一支由科学家、工程师和医学专家组成的跨学科团队， 了解生物学的医生，以及对感染因子的免疫反应， 检测方法，生产诊断设备所需的制造方法， 提供实时响应的流行病学，以及验证诊断设备所需的过程 在现场条件下。我们在探索/试验（R21）阶段的目标是开发一个原型， 基于二维矩阵条形码的机器可读诊断，用于检测血液或血清中的病原体。的 原型将提供1）通过检测循环中的四种不同登革热病毒血清型， 病毒蛋白NS1，以及2）通过检测循环中的埃博拉病毒和登革热病毒的间接鉴定 针对病毒蛋白的抗体。基于现场的可行性测试收集的存档患者血清样本 从登革病毒感染的个体中分离的抗体将提供对 原型化并启用功效计算，以确定R33之前所需的患者数量 发展阶段。对于开发阶段（R33），我们的目标是扩展MMDx器械 用于在护理点分析中检测另外的生物防御剂感染的多重能力。充分 MMDx诊断和实时流行病学的潜力将在一个为期一个月的基于医院的 在登革病毒感染高发的哥伦比亚布卡拉曼加进行现场试验。

7. Project Summary/Abstract This proposal describes the design, construction, and testing of a rapid multiplexed matrix code diagnostic for select biodefense and emerging pathogens. The threat of biological attacks underscores a critical need for rapid point-of care diagnostics that identify specific pathogens in infected persons. Infected or at-risk individuals may be far from critical-care facilities; therefore, diagnostic tests must be easily transportable, stable in storage, simple to use, and easy to interpret. Lateral flow tests (also called immunochromatographic tests or strip tests) are a well-established diagnostic platform because of their simplicity, rapid readout, and low cost qualitative/semi-quantitative detection of antibodies or antigens. Nonetheless, the lack of available point- of-care diagnostics for infectious diseases represents vulnerability in biodefense preparedness as well as a gap in serving public health needs. We propose novel diagnostic device designs, including quick response (QR) two-dimensional matrix-coded readouts, along with attention to chemical coupling approaches, to increase the sensitivity and broaden the multiplex capabilities of lateral flow devices. Effective public health responses also require knowledge of disease distribution and movement based on diagnostic data. Therefore, our proposed Multiplexed Matrix Diagnostic (MMDx) device will be machine-readable, representing a field diagnostic with integrated capability for real time epidemiology using mobile phone technologies and geographic positioning. We have assembled an interdisciplinary team of scientists, engineers, and medical doctors who understand the biology of, and immune responses to infectious agents, the physical chemistry of detection methods, the fabrication methods required to produce diagnostic devices, the technology for providing real-time responsive epidemiology, and the processes required for validating a diagnostic device under field conditions. Our goals for the exploratory/pilot (R21) phase are to develop a prototype two- dimensional matrix barcode-based, machine-readable diagnostic to detect pathogens in blood or serum. The prototype will offer 1) direct identification of the four different dengue virus serotypes by detecting circulating viral protein NS1, as well as 2) indirect Ebola virus and dengue virus identification by detecting circulating antibodies against the viral proteins. A field-based feasibility test archival patient serum samples collected from dengue virus-infected individuals will provide initial validation of the specificity and sensitivity of the prototype and enable power calculations to determine numbers of required patients prior to the R33 developmental phase. For the Developmental Phase (R33), our goals will be to extend the MMDx device multiplex capabilities for detecting additional biodefense agent infections in a point-of-care analysis. The full potential of the MMDx diagnostic and real-time epidemiology will be evaluated in a one-month hospital-based field test in Bucaramanga, Colombia, which has a high incidence of dengue virus infections.