Innovative Approach to Assay Development for the Diagnosis of Invasive Aspergillo

描述(由申请人提供)：侵袭性曲霉病(IA)是免疫功能低下患者中一种毁灭性的、经常是致命的感染，部分原因是早期检测工具不佳。对于曲霉病和其他侵袭性真菌疾病，迅速开展抗真菌治疗的能力对于积极的患者结果至关重要。然而，早期检测的复杂性在于发出疾病开始信号的抗原性标志物水平较低，以及当前诊断分析的特异性较差。这项建议的目的是开发一个基于抗体的诊断平台，由新的单抗与表面增强拉曼光谱和金纳米颗粒夹心分析的超灵敏检测能力相结合。为了应对IA早期检测方面的挑战，我们组建了一个由犹他大学和北达科他州科学家组成的合作研究小组 州立大学由在单克隆抗体开发和开发方面具有专长的免疫学家组成 曲霉菌的筛选；在基础真菌学、真菌化验方面有专长的医学微生物学家 这些专家包括：专注于创新、超灵敏和选择性诊断测试的分析化学家；精通以患者为中心的研究设计和实施的传染病临床医生。我们将分两个阶段阐述这一项目的具体目标。该项目R21阶段的目标是：(1)通过从曲霉属独特的、阶段特定的生物标记物中开发单抗来识别基于蛋白质的真菌靶标。(2)用基于表面增强拉曼光谱的夹心免疫分析技术评价单抗组合。(3)使用不同的人类样本矩阵定义检测性能指标比较。在达到项目第一阶段的里程碑后，R33阶段的目标是：(1)基于生长阶段特异性来表征真菌靶蛋白。(2)优化分析方案，最大限度地提高真菌检测水平。(3)标准化用于曲霉病早期诊断的独特分析试剂盒的组件。通过该项目的成功完成创建的检测战略将通过提高敏感性和特异性重新定义IA的诊断，允许在疾病的最早阶段进行治疗。

DESCRIPTION (provided by applicant): Invasive Aspergillosis (IA) is a devastating and frequently fatal infection in immunocompromised patients, in part because of poor early detection tools. The ability to promptly launch antifungal therapy for Aspergillosis and other invasive fungal diseases is critical for positive patient outcomes. However, early detection is complicated by low levels of antigenic markers that signal disease onset and by poor specificity of current diagnostic assays. The aim of this proposal is to develop an antibody-based diagnostics platform comprised of new monoclonal antibodies coupled with the ultrasensitive detection capabilities of surface enhanced Raman spectroscopy and gold nanoparticle sandwich assays. To address the challenges in the early detection of IA, we have assembled a collaborative research team of scientists from the University of Utah and North Dakota State University composed of immunologists with expertise in monoclonal antibody development and screening for Aspergillus spp; medical microbiologists with strengths in basic mycology, fungal assay development and validation; analytical chemists with a strong focus in the creation of novel, ultrasensitive and selective diagnostic tests; and infectious disease clinicians with proficiency in the design and implementation of patient-oriented research. We will address the specific aims of this project in two phases. The aims for the R21 phase of the project are: (1) Identify protein-based fungal targets through the development of monoclonal antibodies from unique, stage-specific biomarkers of Aspergillus spp. (2) Evaluate monoclonal antibody combinations with surface enhanced Raman spectroscopy-based sandwich immunoassay techniques. (3) Define assay performance metric comparisons using diverse human sample matrices. Upon meeting the milestones of the first stage of the project, the aims of the R33 phase are: (1) Characterize fungal target proteins based on growth stage specificity. (2) Optimize analysis protocols to maximize fungal detection. (3) Standardize components of a unique analysis kit for early clinical Aspergillosis diagnosis. The detection strategy created through the successful completion of this project will redefine the diagnosis of IA through augmented sensitivity and specificity, permitting treatment at the earliest stages of disease.