Innovative Assay Development for the Diagnosis of Invasive Aspergillosis

诊断侵袭性曲霉病的创新检测方法开发

• 批准号: 8032133
• 负责人: Kimberly E. Hanson
• 金额: $ 22.9万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-12-14 至 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8032133
• 关键词: Address; Adsorption; Analytical Chemistry; Antibodies; Antifungal Agents; Antifungal Therapy; Antigens; Applications Grants; Aspergillosis; Aspergillus; Aspergillus fumigatus; Binding; Biological; Biological Assay; Biological Markers; Body Fluids; Breathing; Bronchoalveolar Lavage; Clinical; Clinical Management; Communicable Diseases; Coupled; Data; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Disease; Early Diagnosis; Equipment; Evolution; Fungal Antigens; Generations; Germination; Goals; Gold; Growth; Healthcare Systems; Hospitalization; Human Resources; Immunoassay; Immunocompromised Host; Immunologist; Individual; Infection; Infectious Disease Immunology; Invaded; Label; Laboratories; Medical; Modality; Molds; Monoclonal Antibodies; Morbidity - disease rate; Mus; Nanotechnology; North Dakota; Onset of illness; Outcome; Patients; Performance; Phosphate Buffer; Procedures; Production; Raman Spectrum Analysis; Reproduction spores; Research; Rest; Risk; Saline; Sampling; Scientist; Screening procedure; Sensitivity and Specificity; Serum; Signal Transduction; Silicon Dioxide; Specificity; Specimen; Staging; Surface; Technical Expertise; Testing; The science of Mycology; Time; Tissues; Treatment Protocols; Universities; Urine; Utah; Validation; Work; assay development; attributable mortality; base; clinical Diagnosis; cost; cost effective; design; disease diagnosis; immunosuppressed; innovation; killings; mRNA Differential Displays; man; member; mortality; nanoparticle; novel; patient oriented research; prognostic; research study; tool

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. It presently kills 52% of those infected. 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 the 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, a collaborative research team of scientists from the University of Utah and North Dakota State University has been assembled and includes immunologists with expertise in monoclonal antibody development and screening for Aspergillus spp; medical microbiologists with strengths in basic mycology, and in fungal assay development and validation; analytical chemists and nanotechnologists 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. This R21 grant application will advance this concept using Aspergillus fumigatus (A. fumigatus) - the most common cause of IA - as the proof-of-principle target. The specific aims for this R21 grant application are: (1) To generate unique, stage-specific biomarkers and associated monoclonal antibodies (mAbs) for A. fumigatus. Murine mAbs will be generated against cellular components and secreted products of A. fumigatus. (2) To identify and optimize multiplexed mAb combinations for a sandwich immunoassay based on surface enhanced Raman scattering (SERS), and mAb-coated gold nanoparticle labels. (3) To assess the performance of the assay using biological sample matrices. 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. PUBLIC HEALTH RELEVANCE: Invasive aspergillosis (IA) is a devastating, frequently fatal infection in immunocompromised and immunosup- pressed patients. Unfortunately, the ability to promptly launch antifungal therapy for IA and other invasive fun- gal diseases is compromised by the poor clinically sensitivity, clinical specificity, and prognostic value of exist- ing diagnostic tools. Until IA can be reliably identified at early stages, the morbidity/mortality attendant with this dreadful disease will remain unacceptably high. The overall goal of this project is to develop a rapid, cost effec- tive laboratory test that will serve as a cornerstone for the confident early diagnosis of IA.

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。目前，它杀死了52%的感染者。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。为了解决IA早期检测的挑战，来自犹他州大学和北达科他州州立大学的科学家组成了一个合作研究小组，包括具有单克隆抗体开发和曲霉属筛选专业知识的免疫学家;具有基础真菌学以及真菌测定开发和验证优势的医学微生物学家;分析化学家和纳米技术专家，专注于创造新颖，超灵敏和选择性诊断测试;以及传染病临床医生，精通以患者为导向的研究的设计和实施。 这项R21赠款申请将推进这一概念，使用烟曲霉（A。烟曲霉）-IA的最常见的原因-作为原理证明目标。这项R21资助申请的具体目标是：（1）产生独特的，阶段特异性的生物标志物和相关的单克隆抗体（mAb）。 烟熏。将针对A的细胞组分和分泌产物产生鼠单克隆抗体。 烟熏。 (2)鉴定和优化用于基于表面增强拉曼散射（Sers）和mAb包被的金纳米颗粒标记的夹心免疫测定的多重mAb组合。 (3)使用生物样品基质评估测定法的性能。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。 公共卫生关系：侵袭性曲霉菌病是一种严重的、经常致死的免疫功能低下和免疫抑制患者的感染.不幸的是，现有诊断工具的临床敏感性、临床特异性和预后价值较差，影响了对IA和其他侵袭性真菌疾病及时进行抗真菌治疗的能力。在IA能够在早期阶段被可靠地识别之前，伴随这种可怕疾病的发病率/死亡率将保持不可接受的高水平。该项目的总体目标是开发一种快速、经济有效的实验室检测方法，作为IA早期诊断的基石。