Novel nanoparticular diagnostics for cerebral toxoplasmosis and Chagas in HIV patients living in Latin America

项目摘要 与人类免疫缺陷病毒（HIV）相关的最具破坏性的机会性疾病包括 影响中枢神经系统（CNS）的疾病。它们的非特异性表现使诊断困难， 在资源最充足的环境中，以及在低收入和中等收入国家（LMICs）， 几乎不可能在这种情况下，中枢神经系统感染，如弓形虫脑炎（TE）和中枢神经系统恰加斯病， 疾病通常是凭经验治疗的，当医生的最佳猜测不正确时，这可能是致命的。 据估计，拉丁美洲的弓形虫血清阳性率最高，但艾滋病毒阳性者的诊断率最高。 如果没有昂贵的技术，病人很难接受，而这些技术在资源贫乏的国家是无法获得的。同样地， 查加斯病由克氏锥虫引起，是一种严重的机会性感染， 艾滋病毒/艾滋病，也很难诊断。最后，结核性脑膜炎（TBM）很难诊断， 由于诊断的延误，常常是致命的。33%的秘鲁人和玻利维亚人面临发展风险 在结核病中，当患者是艾滋病毒阳性时，他们有患结核分枝杆菌的风险。南美洲的大部分地区，包括我们的研究地点 在玻利维亚，这两种T. cruzi和T.以及其他常见的中枢神经系统机会性感染 病原体我们提出了一种新的诊断方法的开发和试点测试及其在临床上的应用， 通过以下目的诊断TE和CNS恰加斯病： 具体目标1：开发TE、TBM和CNS查加斯病的平行侧流测定。除了 我们已经开发了查加斯病和结核病的斑点印迹分析，我们有初步的数据表明， 纳米颗粒捕获和检测多个T.弓形虫抗原(3-5)随着进一步发展，我们将 能够将这些单独的免疫测定转换为同时的侧向流测试。 具体目标2。为了确定TE的纳米颗粒诊断测定的灵敏度和特异性， 一个特征明确的HIV感染急性期患者队列中的TBM和CNS恰加斯病 神经系统综合征我们预计弓形虫病，结核性脑膜炎， 隐球菌性脑膜炎的发病率将与其他南美艾滋病毒感染人群相似。我们预计， 然而，CNS恰加斯病也会引起大部分神经系统疾病， 玻利维亚人口，这可能是诊断不足。 该项目将有助于确定拉丁美洲艾滋病毒相关神经系统疾病的临床谱， 使用创新的诊断技术来区分CNS恰加斯病，TBM，TE和其他CNS 感染.当多种病原体流行并迅速致命时， 可以加速启动特异性和潜在的挽救生命的治疗。

PROJECT SUMMARY Among the most devastating opportunistic diseases associated with human immunodeficiency virus (HIV) are those that affect the central nervous system (CNS). Their nonspecific presentation makes diagnosis difficult even in the best-resourced settings, and in Low and Middle Income Countries (LMICs), a definitive diagnosis can be all but impossible. In this context, CNS infections such as toxoplasmic encephalitis (TE) and CNS Chagas disease are often treated empirically, which can be fatal when physicians' best guesses are incorrect. Toxoplasma gondii seroprevalence is estimated to be highest in Latin America, but diagnosis in HIV-positive patients is difficult without expensive technologies that are unavailable in resource-poor countries. Similarly, Chagas disease, caused by Trypanosoma cruzi, is a serious opportunistic infection in people living with HIV/AIDS, and is also difficult to diagnose. Finally, tuberculosis meningitis (TBM) is difficult to diagnose and frequently fatal because of the delay in diagnosis. 33% of Peruvians and Bolivians are at risk for the development of TB, when patients are HIV positive they are at risk of TBM. Much of South America, including our study site in Bolivia, is highly endemic for both T. cruzi and T. gondii as well as other common CNS opportunistic pathogens. We propose the development and pilot testing of a novel diagnostic assays and their use in clinical diagnosis of TE and CNS Chagas disease through the following aims: Specific Aim 1: Development of parallel lateral flow assays for TE, TBM, and CNS Chagas. In addition to our already developed dot blot assays for Chagas and TB, we have preliminary data that demonstrates nanoparticles' ability to capture and detect multiple T. gondii antigens. (3-5) With further development, we will be able to transition these individual immunoassays into a simultaneous lateral flow test. Specific Aim 2. To determine the sensitivity and specificity of the nanoparticle diagnostic assays for TE, TBM and CNS Chagas disease in a well-characterized cohort of HIV-infected patients with acute neurological syndromes. We expect that the proportion of neurological syndromes due to toxoplasmosis, TBM, and cryptococcal meningitis will be similar to those in other South American HIV populations. We anticipate, however, that CNS Chagas disease will also cause a substantial proportion of neurological disease in the Bolivian population, which may have been underdiagnosed. This project will help define the clinical spectrum of HIV-associated neurological disease in Latin America and use innovative diagnostic techniques to distinguish between CNS Chagas disease, TBM, TE, and other CNS infections. Where multiple pathogens are endemic and are rapidly fatal, more rapid, sensitive, and specific tests could accelerate the initiation of specific and potentially life-saving therapy.