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

项目摘要 尽管设计用于方便家庭测试，但快速诊断测试(RDT)通常用于医院- 基于诊断点(POC)的设置，用于诊断寄生虫密度高的有症状患者(&gt；2000 每微升血液)。这在一定程度上是由于当前的横向流动缺乏敏感性和稳定性 RDT设备。在本项目中，我们提出了开发一种新颖的按需纸质诊断平台 这将实现自检(或现场分析)，然后使用便携式远程和集中式信号检测 质谱仪。这种诊断方法是创新的，因为它结合了新级别的简单性和 实用性、适度的成本水平和集中化的检测战略，这将重新定义 用于准确检测疟疾的应用和性价比。更重要的是，我们提出的方法 将提供一个在疾病变得临床明显之前诊断无症状患者的机会， 因此提供负担得起的基于社区的监视和PoC测试。我们将实现这一目标 通过合理设计用于偶联特定抗体的新型离子探针，这将随后 实现使用垂直流体流动格式的纸基免疫分析平台。相比较 有了酶，这些稳定的离子探针就有可能中断、存储和恢复 免疫分析测试，允许在稍后方便的时间进行检测。另外，离子探头也是合理的。 被设计成在刺激下产生小质量的离子，从而允许在野外使用便携式仪器 分析。通过针对所有三个级别的疟疾护理(即，有症状的POC患者、基于监测的 疫情中的无症状检测和现场分析)，我们将提供更大的机会来限制 减少1.88亿疟疾病例，减少每年120亿美元的工人生产力损失。目标1是 设计和合成活性(可裂解)离子探针，作为我们建议的质谱学的质量报告器 基于(MS)的免疫分析技术。还提出了两种信号放大策略来实现 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 纸质MS免疫分析平台实现两个相关任务：离子探针的建立 普通纸基质的介导性免疫分析及新型芯片上MS检测方法的开发 以便于纸张设备的使用。目标3包括通过Close对方法进行验证 与两位分子生物学家(Kingsley Badu--与Pi无关；Knust，加纳，和 克里斯蒂安·科普弗利博士--加州大学欧文分校公共卫生)。James Odei博士(俄亥俄州临床生物统计学 为了进行适当的数据分析和解释，包括了美国州立大学)。作为这一目标的一部分，我们计划筛选 使用开发的试纸对至少1600名(加纳)恶性疟患者/志愿者进行了调查 设备；我们预计将包括约50%的15岁以下儿童。该方法也将在间日疟原虫中得到验证 从埃塞俄比亚采集疟疾样本(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.