Field-deployable Assay for Differential Diagnosis of Malaria and Viral Febrile Illnesses

 描述(由申请人提供)：西非(主要是几内亚、利比里亚和塞拉利昂)目前的埃博拉疫情为发热性疾病诊断的重要性提供了一个宝贵的案例研究。在广泛了解埃博拉疫情之前，出现发烧和寒战的患者可能会纯粹根据症状而被诊断为疟疾，而没有后续测试。如果这种诊断不正确，患者就得不到适当的治疗，如果患者实际上感染了埃博拉或其他传染性病毒，这可能会导致病毒的传播，从而使感染控制复杂化。我们建议开发一种简单、廉价、便携的诊断设备来检测一组可能在西非流行的病毒：埃博拉病毒、拉萨病毒、黄热病病毒、基孔肯雅病毒、登革病毒和西尼罗河病毒；以及恶性疟原虫(人类疟疾最严重的原因)。该设备将非常简单，可以在简单的野战诊所、分诊中心、机场或其他资源匮乏的地点操作，使用带有稳定试剂的新型试剂盒，以消除对冷链的依赖。配套硬件将由电池或太阳能供电，并使用智能手机摄像头进行化验检测、数据分析和报告。该分析基于病原体RNA的等温放大，使用一种新的化学方法，允许使用荧光探针和猝灭剂对目标进行一步、闭管多路检测。这项拟议的工作是桑迪亚国家实验室的生物技术和生物工程小组与德克萨斯大学医学分校的人类健康和免疫研究所合作开展的。拟议的工作从概念验证(R21)阶段开始，在此阶段测试和验证检测和原型硬件对病原体模式菌株的敏感性和特异性。如果成功，这项工作将进入开发阶段(R33)，在此阶段，原型分析和硬件得到改进，并通过临床标本和动物模型感染血液的测试进行验证。

 DESCRIPTION (provided by applicant): The ongoing Ebola epidemic in West Africa (primarily Guinea, Liberia, and Sierra Leone) provides a valuable case study in the importance of diagnostics for febrile illness. Prior to widespread knowledge of the Ebola outbreak, a patient presenting with fever and chills might be diagnosed with malaria, purely on the basis of symptoms with no follow-up tests. If this diagnosis is incorrect, the patient does not get the appropriate therapy, and if the patient in fact has Ebola or other contagious virus, this can lead to spread of the virus which complicates infection control. We propose to develop a simple, inexpensive, portable diagnostic device to detect a panel of viruses that may be endemic in West Africa: Ebola virus, Lassa virus, Yellow Fever virus, Chikungunya virus, Dengue virus, and West Nile virus; as well as Plasmodium falciparum (the most serious cause of human malaria). The device will be simple enough to operate at a simple field clinic, triage center, airport, or other resource-poor location, utilizing a novel assay cartridge with stabilized reagents to eliminate reliance on cold chain. Supporting hardware will be battery- or solar-powered, and uses a smart phone camera for assay detection, data analysis and reporting. The assay is based on isothermal amplification of pathogen RNA, using a novel chemistry that allows single-step, closed tube multiplex detection of targets using fluorescent probes and quenchers. The proposed work is a collaborative effort between the Biotechnology and Bioengineering group at Sandia National Laboratories, and the Institute for Human Health and Immunity at the University of Texas Medical Branch. The proposed work begins with proof-of-concept (R21) phase, during which the assays and prototype hardware are tested and verified for sensitivity and specificity for model strains of pathogens. If successful, the work proceeds to a development (R33) phase, during which the prototype assay and hardware are refined, and validated by testing with clinical specimens and blood from animal model infections.