Metabolomics-based discovery of small molecule biomarkers for noninvasive dengue

项目总结/摘要 流行性登革热（DF）和登革出血热（DHF/DSS）已在整个热带地区出现， 对世界公共卫生造成毁灭性后果。严重登革热（DEN）的急剧增加， 拉丁美洲在过去十年中受到严重关注; 30%的病例现在被诊断为严重登革热 病毒（DENV）感染。在许多发展中国家，DEN是压倒性的临床护理公共卫生能力。 世界该提案的总体目标是为以下疾病的诊断和预后（D&P）提供重大改变： DENV感染。代谢组学方法将用于鉴定候选代谢物小分子 在血清和非侵入性临床标本（尿液和唾液）中均存在的生物标志物（SMB）， 诊断DENV感染并预测进展为严重疾病。使用急性期的初步研究 来自DEN患者的标本已经确定了DF的许多分子特征和候选SMB， 使用基于液相色谱-质谱法（LC-MS）测定血清、唾液和尿液中的DHF/SS 代谢组学在该项目的R21阶段，我们将使用代谢指纹法来确认 在回顾性采集的血清标本中， 在尼加拉瓜进行的高质量儿科DEN医院和队列研究将开始，以描述SMB的特征 以及所涉及的代谢途径，将研究这些SMB在前瞻性收集的 血清，唾液和尿液标本在医院的研究，并将确定一个投资组合的最重要的 区分DEN、严重DEN和非DEN疾病的分子特征。我们将开发算法 包括SMB、临床体征和症状以及DENV感染的D&P的临床实验室结果。在 在项目的R33阶段，前瞻性地从两家医院收集血清、唾液和尿液样本， 研究和基于社区的队列研究将通过使用LC-串联MS（LC-MS）的代谢谱分析进行分析。 MS/MS）来识别候选SMB。选定中小企业的第一代EIA测试将包括在 尼加拉瓜的诊断方案候选人的诊断和预后敏感性和特异性 将确定SMB和“第一代”SMB检测以及SMB的首选临床标本- 基于诊断。总体而言，这些研究将确定一组中小型企业（例如，5-10），将用于 为诊所和医院使用的快速护理点（POC）测试制定目标产品简介（TPP） 检测唾液和尿液中的SMB，预测严重DEN是创新的， 提供了一个真正的诊断范式转变的机会，通过使用廉价，易于采购，非 用于DEN的D&P的侵袭性临床标本。

PROJECT SUMMARY / ABSTRACT Epidemic dengue fever (DF) and dengue hemorrhagic fever (DHF/DSS) have emerged throughout the tropical world with devastating public health consequences. A dramatic increase in severe dengue disease (DEN) in Latin America in the last decade is of grave concern; 30% of cases are now diagnosed as severe dengue virus (DENV) infections. DEN is overwhelming public health capacity for clinical care in much of the developing world. The overall goal of this proposal is to provide a major change in the diagnosis and prognosis (D&P) of DENV infections. A metabolomics approach will be used to identify candidate metabolite small molecule biomarkers (SMBs) that occur both in serum and in non-invasive clinical specimens (urine and saliva) that diagnose DENV infection and predict progression to severe disease. Preliminary studies using acute phase specimens from DEN patients have identified a number of molecular features and candidate SMBs of DF and DHF/SS in serum, saliva, and urine using liquid chromatography-mass spectrometry (LC-MS)-based metabolomics. In the R21 phase of this project, we will use a metabolic fingerprinting approach to confim existing and identify new candidate SMBs in retrospectively collected serum specimens available from the high-quality pediatric DEN hospital-based and cohort studies in Nicaragua, will begin to characterize the SMBs and metabolic pathways involved, will investigate the efficacy for D&P of these SMBs in prospectively collected serum, saliva, and urine specimens in the hospital study, and will identify a portfolio of the most significant molecular features that differentiate DEN, severe DEN, and non-DEN disease. We will develop algorithms including SMBs, clinical signs and symptoms, and clinical laboratory results for the D&P of DENV infections. In the R33 phase of the project, prospectively collected serum, saliva, and urine samples from both the hospital study and a community-based cohort study will be analyzed by metabolic profiling using LC-tandem MS (LC- MS/MS) to identify candidate SMBs. First generation EIA tests for selected SMBs will be included in the diagnostic regimen in Nicaragua The diagnostic and prognostic sensitivity and specificity of the candidate SMBs and "first generation" SMB tests will be determined as will the preferred clinical specimen for SMB- based diagnoses. Overall these studies will identify a panel of SMBs (e.g., 5-10), which will be used to formulate the Target Product Profiles (TPP) for rapid point-of-care (POC) tests for use in clinics and hospitals for DEN D&P. Detection of SMBs in saliva and urine that are predictive of severe DEN is innovative and provides the opportunity for a true paradigm shift in diagnosis by using inexpensive, easily procured, non- invasive clinical specimens for D&P of DEN.