Simultaneous Diagnosis of Dengue and Prognosis of Severe Dengue with a Single Point-of-Care Test

登革热是最常见的由蚊子传播的病毒性疾病，导致极大的发病率和死亡率 和经济负担。每年，估计有3.9亿(M)登革病毒(DENV)感染，9600万 有症状的登革热病例和50万例重度登革热病例。未经治疗的严重登革热 病死率为20%-40%，但如果及时住院治疗，病死率降至1%以下。不幸的是， 严重的登革热现在只有通过熟练地观察临床警告信号才能被发现，并进展为严重的 疾病可能在几个小时内发生，使有效的护理变得困难。因此，迫切需要 可以识别最有可能进展为严重登革热的患者的预后测试。有四种血清型 任何一种血清型的DENV感染都会产生强大的、潜在的终身免疫力，通过 相同的血清型，这是由于针对血清型特异性表位的中和抗体的发展。 然而，首次感染引起的大多数抗体是交叉反应和非中和的，而且矛盾的是 会增加后来感染不同血清型的患者患严重登革热的风险，原因是 一种称为抗体依赖增强(ADE)的现象。异型继发感染是指 严重登革热的主要公认危险因素，但目前无法确定异型继发感染 在可供决策的时间内影响临床护理和结果。 我们建议创建一种简单、廉价、具有可视加/减读数的关注点(POC)测试 这不仅可以早期诊断登革热病毒感染，而且可以迅速预测严重登革热的预后。 识别有继发性异型感染的患者。 第一阶段将创建关键的生物标记物检测试剂，并将它们合并到两套特定于血清类型的 横向流动分析(LFAs)：1)抗原LFAs用于诊断和分型当前感染(通过检测DENV- 在活动性感染期间制作的NS1)，以及2)用于诊断和分型既往感染的抗体LFA(通过 检测患者对以前的感染反应产生的抗DENV抗体)。第二阶段将结合 单个分析在单个双报告LFA中进行，然后生成分析和临床性能数据 支持CE标志和FDA 510(K)认证申请成为POC IVD。 创新：该方案创造性地结合了成熟的临床观察(异型次要 感染是严重登革热的主要危险因素)，有两个重大的技术进步：1)快速血清分型 以及2)过去感染的快速血清分型。有了这个组合，我们将开发一款第一流的- 将改变临床实践的DENV诊断和严重登革热预后的类别POC试验。 影响：一种简单、廉价的POC测试，即使在资源有限的情况下也能提供及时的预后结果 环境将使登革热发病率和死亡率大幅下降，同时降低医疗成本 通过允许临床医生有效和高效地将资源集中在可能最需要它们的患者身上。

Dengue is the most common mosquito-borne viral disease and the cause of tremendous morbidity, mortality and economic burden. Yearly, there are an estimated 390 million (M) Dengue virus (DENV) infections, 96M symptomatic cases of Dengue Fever, and 0.5M cases of Severe Dengue. Untreated Severe Dengue has a fatality rate of 20-40%, but the fatality rate drops to less than 1% with timely in-hospital care. Unfortunately, Severe Dengue is now detected only by skillful observation of clinical warning signs, and progression to severe disease can occur within hours, making effective care difficult. Therefore, there is an urgent need for prognostic tests that can identify patients most likely to progress to Severe Dengue. There are four serotypes of DENV and infection with any one serotype generates strong, potentially life-long immunity to reinfection by that same serotype, due to development of neutralizing antibodies against serotype-specific epitopes. However, most antibodies induced by first infections are cross-reactive and non-neutralizing, and paradoxically can increase the risk of Severe Dengue in patients who suffer a later infection with a different serotype, due to a phenomenon termed Antibody-Dependent Enhancement (ADE). Heterotypic secondary infection is the major accepted risk factor for Severe Dengue but heterotypic secondary infections cannot now be identified within the time available for decision making to impact clinical care and outcome. We propose to create a simple, inexpensive, Point-of-Care (POC) test with visual plus/minus readout that will not only diagnose DENV infections early, but also be prognostic for Severe Dengue by quickly identifying patients who have secondary heterotypic infections. Phase I will create key biomarker detection reagents and incorporate them into two sets of serotype-specific Lateral Flow Assays (LFAs): 1) antigen LFAs to diagnose and serotype current infections (by detecting DENV- NS1 made during active infections), and 2) antibody LFAs to diagnose and serotype past infections (by detecting anti-DENV-antibodies made by patients in response to previous infections). Phase II will combine the individual assays in a single dual reporter LFA and then generate analytical and clinical performance data to support applications for CE Mark and FDA 510(k) clearance as a POC IVD. Innovation: The proposal creatively combines a well-established clinical observation (heterotypic secondary infection is the major risk factor for Severe Dengue), with two significant technical advances:1) rapid serotyping of current infections, and 2) rapid serotyping of past infections. With this combination we will develop a first-in- class POC test for DENV diagnosis and Severe Dengue prognosis that will change clinical practice. Impact: A simple, inexpensive POC test that provides timely prognostic results even in resource-limited settings would enable dramatic reductions in Dengue morbidity and mortality while reducing health care costs by allowing clinicians to focus resources effectively and efficiently on patients who may need them most.