Innovative Approach to Assay Development for the Diagnosis of Invasive Aspergillo

描述（由申请人提供）：侵袭性支气管炎（IA）是免疫功能低下患者的一种破坏性和经常致命的感染，部分原因是早期检测工具不佳。 及时启动抗真菌治疗曲霉菌病和其他侵袭性真菌病的能力对于患者的积极结局至关重要。然而，早期检测是复杂的低水平的抗原标志物，信号疾病发作和目前的诊断测定的特异性差。该提案的目的是开发一种基于抗体的诊断平台，该平台由新的单克隆抗体与表面增强拉曼光谱和金纳米颗粒夹心测定的超灵敏检测能力相结合组成。 为了应对IA早期检测的挑战，我们组建了一个由来自犹他州和北达科他州大学的科学家组成的合作研究团队 州立大学由具有单克隆抗体开发专业知识的免疫学家组成， 筛选曲霉属;医学微生物学家，在基础真菌学、真菌测定方面具有优势 开发和验证;分析化学家专注于创造新颖，超灵敏和选择性诊断测试;传染病临床医生精通以患者为导向的研究的设计和实施。我们将分两个阶段实现该项目的具体目标。该项目的R21阶段的目标是：（1）通过从曲霉属的独特的阶段特异性生物标志物开发单克隆抗体来鉴定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测定技术评价单克隆抗体组合。(3)使用不同的人体样本基质定义测定性能指标比较。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。(3)标准化用于早期临床曲霉病诊断的独特分析试剂盒的组分。通过成功完成该项目创建的检测策略将通过增强的灵敏度和特异性重新定义IA的诊断，从而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.