Metabolomics-based discovery of small molecule biomarkers for noninvasive dengue

项目摘要/摘要 在整个热带地区出现了流行性登革热和登革出血热。 给世界带来了毁灭性的公共卫生后果。年严重登革热(DEN)病例急剧增加 拉丁美洲过去十年的情况令人严重关切；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在临床护理方面的公共卫生能力是压倒性的 世界。这项建议的总体目标是在诊断和预后(D&P)方面提供重大变化 DENV感染。将使用代谢组学方法来鉴定候选代谢物小分子 出现在血清和非侵入性临床标本(尿液和唾液)中的生物标记物(SMB) 诊断DENV感染并预测病情发展为严重疾病。急性期的初步研究 来自DEN患者的样本已经确定了一些DF和SMB的分子特征和候选SMB 液质联用法测定血清、唾液和尿液中的DHF/SS 代谢组学。在此项目的R21阶段，我们将使用代谢指纹方法来配置 在追溯收集的血清样本中确定现有的SMB并确定新的候选SMB 尼加拉瓜的高质量儿科医院研究和队列研究将开始成为中小型企业的特征 以及涉及的代谢途径，将调查这些SMB在前瞻性收集中对D&P的疗效 在医院研究的血清、唾液和尿液样本，并将确定最重要的组合 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DENV感染的SMBS、临床体征和症状以及临床实验室结果。在……里面 项目的R33阶段，预期从两家医院收集血清、唾液和尿液样本 这项研究和一项以社区为基础的队列研究将通过使用LC-Tandem MS(LC-Tandem MS)进行代谢谱分析 MS/MS)以确定候选中小型企业。选定中小型企业的第一代环评测试将包括在 尼加拉瓜的诊断方案：候选患者诊断和预后的敏感性和特异性 SMB和第一代SMB测试将被确定，SMB的首选临床标本将被确定- 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家)，用于 制定用于诊所和医院的快速护理点(POC)测试的目标产品配置文件(TPP) 对于DEN D&P来说，检测唾液和尿液中可预测严重DEN的SMBS是一项创新和 提供了真正转变诊断模式的机会，方法是使用价格低廉、易于获得、非 DEN 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.