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

描述(由申请人提供)：本提案描述了一种用于选定的生物防御和新出现的病原体的快速多路复用矩阵码诊断的设计、构造和测试。生物攻击的威胁突出表明，迫切需要进行快速护理点诊断，以确定感染者中的特定病原体。感染者或高危个体可能远离重症监护设施；因此，诊断测试必须易于携带， 储存稳定，使用简单，易于解释。横向流动试验(也称为免疫层析试验或试条试验)是一种成熟的诊断平台，因为它们简单、快速读出，并对抗体或抗原进行低成本的定性/半定量检测。尽管如此，缺乏对传染性疾病的可用的看护点诊断 疾病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)二维矩阵编码读数，以及对化学耦合方法的关注，以提高横向流动设备的灵敏度并扩大其多路复用能力。有效的公共卫生对策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，代表着一种现场诊断，具有使用移动电话技术和地理定位进行实时流行病学的集成能力。我们组建了一支由科学家、工程师和医生组成的跨学科团队，他们了解传染病病原体的生物学和免疫反应、检测方法的物理化学、制造诊断设备所需的制造方法、提供实时响应流行病学的技术，以及在现场条件下验证诊断设备所需的流程。我们在探索/试验阶段(R21)的目标是开发一种基于二维矩阵条形码的、机器可读的诊断原型，以检测血液或血清中的病原体。原型机将提供1)直接识别 通过检测循环病毒蛋白NS1以及 2)通过检测循环抗体间接鉴定埃博拉病毒和登革热病毒 病毒蛋白。现场可行性测试存档从登革病毒感染者身上收集的患者血清样本将提供原型的特异性和敏感性的初步验证，并使功率计算能够在R33发育阶段之前确定所需的患者数量。在开发阶段(R33)，我们的目标将是扩展MMDx设备的多路传输能力，用于在护理点分析中检测更多的生物防御剂感染。MMDx诊断和实时流行病学的全部潜力将在登革热病毒感染高发的哥伦比亚布卡拉曼加进行为期一个月的医院现场测试中进行评估。

DESCRIPTION (provided by applicant): 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 sensitivit 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.