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

登革热是最常见的蚊媒病毒性疾病， 经济负担。每年，估计有3.9亿（M）登革病毒（DENV）感染，96 M 有症状的登革热病例和0.5M严重登革热病例。未经治疗的严重登革热 死亡率为20- 40%，但通过及时的住院治疗，死亡率降至1%以下。不幸的是， 现在，只有通过熟练观察临床警告信号，并发展为严重的登革热， 疾病可在数小时内发生，使有效护理变得困难。因此，迫切需要 预后测试可以识别最有可能进展为严重登革热的患者。有四种血清型 感染任何一种血清型，通过以下方式产生对再感染的强大的、潜在的终身免疫力： 由于针对血清型特异性表位的中和抗体的发展，相同的血清型。 然而，首次感染诱导的大多数抗体是交叉反应性和非中和性的， 可能会增加严重登革热的风险，患者谁遭受后来感染不同的血清型，由于 一种称为抗体依赖性增强（ADE）的现象。异型继发感染是指 严重登革热的一个主要公认的危险因素，但现在无法确定异型继发感染 在可用于影响临床护理和结果的决策的时间内。 我们建议创建一个简单，廉价，床旁（POC）测试与视觉加/减读出 这不仅可以早期诊断登革病毒感染，而且可以通过快速诊断来预测重症登革， 识别继发性异型感染的患者。 第一阶段将创建关键的生物标志物检测试剂，并将其纳入两套针对特定类型的 侧流测定（LFA）：1）抗原LFA，用于诊断和血清分型当前感染（通过检测DENV-1）。 在活动性感染期间产生的NS 1），和2）抗体LFA以诊断和血清分型过去的感染（通过 检测由患者响应于先前感染而产生的抗DENV抗体）。二期将联合收割机 单个双报告基因LFA中的单个检测试剂盒，然后生成分析和临床性能数据 支持作为POC IVD申请CE认证和FDA 510（k）许可。 创新：该提案创造性地结合了完善的临床观察（异型继发性 感染是严重登革热的主要危险因素），有两项重大技术进步：1）快速血清分型 当前感染，和2）过去感染的快速血清分型。通过这种组合，我们将开发出一种先进的 用于DENV诊断和严重登革热预后的POC类测试将改变临床实践。 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，同时降低卫生保健成本 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.