Malaria Management through an On-demand Diagnostic Approach using Novel Ionic Probes

项目摘要 尽管快速诊断测试（RDT）被设计为便于家庭测试，但通常在医院中使用- 用于诊断寄生虫密度高（>2000）的症状性患者的基于床旁（POC）的设置 每微升血液）。这在一定程度上是由于目前基于横向流动的方法缺乏灵敏度和稳定性。 RDT设备。在目前的项目中，我们建议开发一种新的按需纸基诊断平台 这将使自我测试（或现场分析），随后是远程和集中的信号检测使用便携式 质谱仪这种诊断方法是创新的，因为它结合了新的简单性水平， 实用性，适度的成本水平，以及集中的检测策略，这将重新定义 准确检测疟疾的应用和性能/成本比。更重要的是，我们提出的方法 将提供在疾病变得临床明显之前诊断无症状患者的机会， 从而提供负担得起的社区监测和POC检测。我们会达到这个目标 通过合理设计用于偶联特异性抗体的新型离子探针， 能够实现利用垂直流体流动形式的纸基免疫测定平台。相比 对于酶，这些稳定的离子探针使中断、存储和恢复酶的能力成为可能。 免疫分析测试，允许在稍后方便的时间检测。此外，离子探针是合理的， 设计为在刺激时产生小质量的离子，允许使用便携式仪器进行现场测试。 分析.通过针对所有三级疟疾护理（即，症状性POC患者，基于监测 无症状的检测和现场分析），我们将提供更大的机会， 1.88亿例疟疾病例，并减少每年120亿美元的工人生产力损失。目标1： 设计和合成活性（可裂解）离子探针作为我们提出的质谱的质量报告 （MS）为基础的免疫测定技术。还提出了两种信号放大策略， 从手指刺血中超灵敏地检测低寄生虫密度。目标2将开发和优化 通过实现两个相互关联的任务：离子探针的建立， 介导的免疫分析在普通纸基板上，和新的芯片上的MS检测方法的发展 以便于纸张装置的使用。目标3涉及方法的验证，通过关闭 与两名分子生物学家（Kingsley Badu -与PI无关; KNUST，加纳和 博士Cristian Koepfli -加州大学欧文分校，公共卫生）。James Odei博士（临床生物统计学，俄亥俄州 州立大学）进行适当的数据分析和解释。作为这一目标的一部分， 至少1，600名恶性疟原虫（P）患者/志愿者（在加纳）使用开发的论文 设备;我们预计将包括约50%的15岁以下儿童。该方法还将针对间日疟原虫进行验证 从埃塞俄比亚收集的疟疾样本（预计约600名志愿者）。

Project Summary Although designed to facilitate home testing, rapid diagnostic test (RDT) have typically been used in a hospital- based point-of-care (POC) setting for diagnosis of symptomatic patients where parasite density is high (>2000 per microliter of blood). This is, in part, due to the lack of sensitivity and stability of current lateral flow-based RDT devices. In the current project, we propose to develop a novel on-demand paper-based diagnostic platform that will enable self-testing (or field analysis) followed by remote and centralized signal detection using portable mass spectrometers. This diagnostic approach is innovative as it combines new levels of simplicity and practicality, modest levels of cost, and a centralized detection strategy, which will redefine the breadth of application and performance/cost ratio for accurate malaria detection. More importantly, our proposed approach will provide an opportunity to diagnose asymptomatic patients before the disease becomes clinically apparent, therefore providing affordable community-based surveillance and POC tests. We will achieve this objective through the rational design of novel ionic probes for coupling to specific antibodies, which will subsequently enable the implementation of a paper-based immunoassay platform utilizing vertical fluid flow format. Compared with enzymes, these stable ionic probes make possible the ability to interrupt, store, and restore the immunoassay test, allowing detection at a later convenient time. In addition, the ionic probes are rationally designed to yield ions of small masses upon stimulation permitting the use of portable instruments for field analysis. By targeting all three levels of malaria care (i.e., symptomatic POC patients, surveillance-based asymptomatic detection and field analysis in an outbreak), we will offer a greater chance of limiting the 188 million cases of malaria and reduce the $12 billion yearly loss in worker productivity. Aim 1 is to design and synthesize active (cleavable) ionic probes as mass reporters for our proposed mass spectrometry (MS)-based immunoassay technology. Two signal amplification strategies are also proposed to enable ultrasensitive detection of low parasite density from a finger prick blood. Aim 2 will develop and optimize the paper-based MS immunoassay platform by implementing two interrelated tasks: establishment of ionic probe- mediated immunoassay on ordinary paper substrate, and the development of new on-chip MS detection methods to facilitate the ease of use of the paper devices. Aim 3 involves validation of the method, through close collaborative interactions with two molecular biologists (Kingsley Badu – not related to PI; KNUST, Ghana, and Dr. Cristian Koepfli - University of California, Irvine, Public Health). Dr. James Odei (Clinical Biostatistics, Ohio State University) is included for proper data analysis and interpretation. As part of this aim, we project screening of at least 1,600 patients/volunteers with Plasmodium (P) falciparum (in Ghana) using the developed paper device; we expect to include ~50% children under age 15. The method will also be validated against P. vivax malaria samples (~600 volunteers expected) collected from Ethiopia.