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

项目总结 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 那些影响中枢神经系统(CNS)的疾病。它们的非特异性表现甚至使诊断变得困难 在资源最充足的情况下，在低收入和中等收入国家(LMIC)，一个明确的诊断可能是 几乎是不可能的。在这方面，弓形体脑炎和查加斯等中枢神经系统感染 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，弓形虫血清阳性率在拉丁美洲最高，但在艾滋病毒阳性的患者中被诊断为弓形虫 如果没有昂贵的技术，患者是很困难的，而这些技术在资源匮乏的国家是无法获得的。同样， 由克氏锥虫引起的恰加斯病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，结核病脑膜炎(TBM)很难诊断和 由于诊断的延迟，常常是致命的。33%的秘鲁人和玻利维亚人面临着发展的风险 在结核病方面，当患者艾滋病毒呈阳性时，他们有患结核病的风险。南美洲大部分地区，包括我们的研究站点 在玻利维亚，克鲁兹弓形虫和弓形虫以及其他常见的中枢神经系统机会性疾病高度流行 病原体。我们建议开发和中试一种新的诊断分析方法，并将其用于临床。 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 Chagas的平行侧向流动分析方法。除了……之外 我们已经开发了查加斯和结核病的斑点印迹分析，我们有初步数据表明 纳米颗粒捕获和检测多种弓形虫抗原的能力。(3-5)随着进一步的发展，我们将 能够将这些单独的免疫分析转换为同步的侧向流动测试。 具体目的2.为了确定用于TE的纳米颗粒诊断分析的敏感性和特异性， 急性HIV感染者队列中的特发性结缔组织病和中枢神经系统恰加斯病 神经综合征。我们预计，由于弓形虫病、TBM、 隐球菌性脑膜炎将与南美其他艾滋病毒人群的脑膜炎相似。我们预计， 然而，中枢神经系统恰加斯病也会导致相当大比例的神经系统疾病 玻利维亚人口，这可能被低估了。 该项目将有助于确定拉丁美洲和拉丁美洲艾滋病毒相关神经疾病的临床谱系 使用创新的诊断技术来区分CNS Chagas病、TBM、TE和其他CNS 感染。如果多种病原体是地方性的，并且是迅速致命的，则进行更快速、更敏感和更特异的检测 可以加速启动特定的和潜在的挽救生命的疗法。

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.