Addressing the Global Burden of Leptospirosis in Two Endemic Countries

 描述(由申请人提供)：拟议的U19 ICIDR计划重点关注钩端螺旋体的人类感染和疾病。我们的总体方法是使用基本的生物科学方法，结合秘鲁和斯里兰卡不同流行病学背景下的临床/现场研究，开发新的、临床上可操作的诊断测试。考虑到钩端螺旋体在当地、区域和国际上的多样性，拟议的研究解决了该领域迄今无法克服的一个空白，即获得廉价、敏感和有效的钩端螺旋体病诊断测试，但无论潜在的传播动态如何，都易于部署和有效。这些测试的准确性和效率的提高将使及时实施适当的治疗成为可能，这对改善临床结果至关重要。本方案的总体目标是开发强有力的工具和方法，有效量化全球钩端螺旋体病的负担，解决世界卫生组织钩端螺旋体病流行病学参考小组(LEG)阐明的国际钩端螺旋体病研究界共同的几个关键目标。1、2这些方法适用于所有钩端螺旋体病流行和流行地点，摘要如下： 1.在秘鲁(项目1)和斯里兰卡(项目2)已知或疑似高度流行的环境中，对疑似急性钩端螺旋体病病例进行前瞻性现场临床研究，这两个国家的钩端螺旋体多样性很高，包括最高致病性以及致病性较低(但仍具有传染性)的钩端螺旋体。这些地点将包括秘鲁和斯里兰卡的城市与农村以及季节性与常年钩端螺旋体病风险地区的对比。这些项目将侧重于住院病例(病情较重)和门诊病人(一般病情较轻)，并致力于全面跟踪临床和微生物/分子结果； 2.用常规血清学(MAT、ELISA法)、3、4、5常规培养、我们发表的非培养非依赖于培养的分子技术(常规PCR、6、7 qPCR、8多位点序列分型(MLST)9、10)鉴定现场研究中的钩端螺旋体病样本； 3.基于我们最新开发的钩端螺旋体基因组计划数据库(PATRIC，http://patricbrc.org；存放在GenBank，http://www.ncbi.nlm.nih.gov/bioproject/?term=leptospira(均于2014年3月7日访问))，使用基于蛋白质微阵列技术的新开发的抗原检测测试(脂多糖；或者，钩端螺旋体蛋白)和基于抗体的血清学测试，与传统诊断(金标准重新定义为MAT(配对样本)加血/尿聚合酶链式反应)进行对照，考虑到当前培养技术的不敏感性(参考文献11，12讨论了过去金标准诊断的问题)。 该计划的预期成果是1)诊断钩端螺旋体病和确定人类钩端螺旋体感染的新工具；2)定义和优化这些新工具在秘鲁和斯里兰卡及其他地区具有代表性的多样化环境中的普及性(例如，通过美国疾病控制和预防中心接收来自世界各地的样本；见Renee Gloway，支持函)；以及3)通过促进独立创造性科学思维的发展、假设的产生和测试以及解决流行国家最优先的临床和翻译研究问题的新方法，提高外国合作者的研究能力。该项目的其他好处包括明确应用于OneHealth药物，因为人类钩端螺旋体病反映了人与人畜共患病的家庭和农业来源的相互作用，如狗、牛、猪和水牛、野生和有袋类啮齿动物和有袋动物。这里开发的工具将直接应用和翻译到兽医和农业环境中，但这些工作将超出本项目的范围。

 DESCRIPTION (provided by applicant): The proposed U19 ICIDR Program is focused on human infection and disease by Leptospira. Our overall approach is to use fundamental biological scientific methods combined with clinical/field studies in contrasting epidemiological contexts in Peru and Sri Lanka, to develop new, clinically actionable diagnostic tests. Taking into account the local, regional, and international diversity of Leptospira, the proposed research addresses a hitherto insurmountable gap in the field, that of obtaining diagnostic tests for leptospirosis that are inexpensive, sensitive and efficient, yet easily deployable and effective regardless of the underlying transmission dynamics. The improved accuracy and efficiency of these tests will enable timely administration of appropriate therapy, which is vital for improving clinical outcomes. The overall goal of this Program is to develop robust tools and approaches that will effectively quantify the Global Burden of Leptospirosis, addressing several key goals shared by the international leptospirosis research community as articulated by the World Health Organization's Leptospirosis Epidemiology Reference Group (LERG).1, 2 These approaches, which are applicable to all leptospirosis-endemic and epidemic-prone sites, are summarized as follows: 1. Carry out prospective, field-based clinical studies of suspected acute leptospirosis cases in known or suspected highly endemic settings in Peru (Project 1) and Sri Lanka (Project 2) where the diversity of Leptospira is high and includes the most highly pathogenic as well as less pathogenic (but still infectious) Leptospira. These sites will include contrasting urban vs. rural and seasonal vs. perennial leptospirosis risk areas in Peru and Sri Lanka. The projects will focus on both hospitalized cases (more severe disease) and outpatients (generally mild spectrum of disease) with dedicated attention to comprehensively follow up clinical and microbiological/molecular outcomes; 2. Characterize serum and urine samples from field studies with regard to leptospirosis diagnosis using conventional serology (MAT, ELISA),3, 4, 5 routine culture, our published culture-independent molecular techniques (conventional PCR,6, 7 qPCR,8 Multi Locus Sequence Typing (MLST)9, 10); 3. Test newly developed antigen-detection tests (LPS; alternatively, Leptospira proteins) and antibody- based serological tests using protein microarray technology based on our newly developed Leptospira Genome Project database (PATRIC, http://patricbrc.org; deposited in GenBank, http://www.ncbi.nlm.nih.gov/bioproject/?term=leptospira (both accessed March 7, 2014)) against conventional diagnostics (gold standard re-defined as MAT (on paired samples) plus PCR of blood/urine, given the insensitivity of current culture techniques (problems with past gold-standard diagnostics discussed in Refs.11, 12). The expected outcomes of this program are 1) New tools to diagnose leptospirosis and to identify leptospiral infection of humans; 2) Define and optimize the generalizability of these new tools in diverse and representative settings in Peru and Sri Lanka, and beyond (for example, through the U.S. Centers for Disease Control and Prevention that receives samples from all over the world; see Renee Galloway, Letter of Support); and 3) Enhancing research capacity of foreign collaborators by facilitating development of independent creative scientific thinking, hypothesis generation and testing, and new approaches to addressing clinical and translational research problems of highest priority to the endemic country. Additional benefits from this project include clear application to OneHealth medicine because human leptospirosis reflects interactions of humans with zoonotic domestic and agricultural sources such as dogs, cattle, pigs, and water buffalo, wild and peridomestic rodents and marsupials. The tools developed here will have direct application and translation to veterinary and agricultural settings but such work will be beyond the scope of the present project.