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

7.项目摘要/摘要 本方案描述了一种快速多路矩阵编码诊断系统的设计、构造和测试 选择生物防御和新出现的病原体。生物攻击的威胁突显出迫切需要 快速护理点诊断，识别感染者中的特定病原体。受感染或处于危险之中 个人可能远离重症监护机构；因此，诊断测试必须易于携带， 储存稳定，使用简单，易于解释。侧向流动试验(也称为免疫层析法 测试或条带测试)是一种成熟的诊断平台，因为它们简单、快速读出和低成本 抗体或抗原的定性/半定量检测成本。尽管如此，缺乏可用的点- 对传染病的护理诊断代表着生物防御准备方面的脆弱性，以及 在满足公共卫生需求方面存在差距。我们提出了新的诊断设备设计，包括快速响应 (QR)二维矩阵编码读数，以及对化学耦合方法的关注，以 提高横向流动装置的灵敏度并扩大其多路传输能力。有效的公共卫生 应对措施还需要根据诊断数据了解疾病分布和运动情况。因此， 我们建议的多路复用矩阵诊断(MMDx)设备将是机器可读的，代表一个领域 使用移动电话技术和集成功能进行实时流行病学诊断 地理定位。我们已经组建了一支由科学家、工程师和医疗人员组成的跨学科团队 了解感染性病原体的生物学和免疫反应的医生 检测方法，制造诊断设备所需的制造方法， 提供实时响应流行病学，以及验证诊断设备所需的流程 在田间条件下。我们在探索/试验阶段(R21)的目标是开发两个原型- 基于维度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。这个 Prototype将提供1)通过检测循环来直接识别四种不同的登革热病毒血清型 病毒蛋白NS1，以及2)通过检测循环间接识别埃博拉病毒和登革热病毒 针对病毒蛋白的抗体。现场采集患者血清标本的可行性检验 将提供对登革热病毒感染个体的特异性和敏感性的初步验证 制作原型并启用功率计算，以确定R33之前所需的患者数量 发展阶段。在开发阶段(R33)，我们的目标是扩展MMDx设备 在护理点分析中检测其他生物防御剂感染的多重能力。完整的 MMDx诊断和实时流行病学的潜力将在一个月的医院基础上进行评估 在登革热病毒感染高发的哥伦比亚布卡拉曼加进行现场检测。

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.