Multiplexed rapid immunoassay for invasive fungal disease

侵袭性真菌病的多重快速免疫分析

• 批准号: 10116147
• 负责人: Thomas R Kozel
• 金额: $ 70.26万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-23 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10116147
• 关键词: Address; Advanced Development; Animal Model; Antibodies; Antifungal Agents; Archives; Aspergillosis; Aspergillus; Biological Assay; Biological Markers; Body Fluids; Cancer Patient; Candida; Candidiasis; Caring; Cell Wall; Clinical; Collaborations; Communicable Diseases; Coupled; Cryptococcal Meningitis; Diagnosis; Diagnostic; Diagnostic tests; Disease; Disseminated candidiasis; Early Diagnosis; Early treatment; Enzyme-Linked Immunosorbent Assay; Epitopes; Evaluation; Florida; Freezing; Fusarium; Goals; Health Care Costs; Hematogenous; Hematopoietic Stem Cell Transplantation; Histoplasma; Human; Immune; Immune response; Immunoassay; Individual; Infection; Lateral; Literature; Mannans; Medical; Medical center; Methods; Molds; Monoclonal Antibodies; Mucorales; Mucormycosis; Mus; Mycoses; Nevada; Organ Transplantation; Pathogenesis; Patient Care; Patient-Focused Outcomes; Patients; Performance; Pharmaceutical Preparations; Plant Roots; Plasma; Polysaccharides; Primary Health Care; Probability; Production; Reporting; Research; Research Project Grants; Sampling; Solid; Specimen; Splenocyte; Technology; The science of Mycology; Translational Research; Transplantation; United States National Institutes of Health; Universities; Urine; Yeasts; antimicrobial; cancer therapy; fungus; galactomannan; health care settings; high risk; human model; improved; monoclonal antibody production; mortality; mouse model; pathogen; patient population; pre-clinical; prospective; prototype; rapid diagnosis; success; user-friendly

Recent advances in i) treatment of cancer patients, ii) hematopoietic stem cell and organ transplantation, iii) use of potent immunosuppressive drugs and iv) advanced medical care have created an expanding population of patients at high risk for invasive fungal diseases (IFD) produced by common environmental molds (aspergillosis, mucormycosis and fusariosis) and normally commensal yeast (candidiasis). Numerous studies have shown that early diagnosis and treatment with the right antifungal agent will improve patient outcome. The goal of this translational research project is a multiplexed immunoassay that will use serum/plasma or urine as a clinical specimen for diagnosis of early-stage infection by Aspergillus spp., Fusarium spp., the Mucorales, and Candida spp. The target patient population will be individuals who are at high risk for fungal infection due to use of immunosuppressive drugs or advanced medical care. The approach will be an immunoassay that detects cell wall mannans of each fungus that are shed into body fluids during infection. The product will be an immunoassay, most likely in the lateral flow immunoassay (LFIA) format, for use at primary health-care settings. The immunoassay will be sensitive, specific, rapid, inexpensive, and user friendly. There are four Specific Aims. Aim 1 will further develop and validate high-quality monoclonal antibodies (mAbs) specific for mannan epitopes that are unique to each of the targeted groups of fungi. Aim 2 will determine the analytical sensitivity of an immunoassay that would be needed to detect early-stage infection using specimens from humans and murine models. Aim 3 will develop multiplexed immunoassays that have the analytical sensitivity needed for early diagnosis. Aim 4 will assess clinical performance of prototype immunoassays using plasma and urine from animal models and patients with IFD. This study addresses a critical unmet need for diagnosis of increasingly common and fatal opportunistic fungal infections that occur in high risk patients. If successful, the multiplexed assay will be a game changer for patient care. The proposal is supported by a strong premise from the literature and solid preliminary results. These preliminary studies, coupled with the existing paradigm for using immunoassays that target shed cell wall mannans for diagnosis of IFD, make the probability for success very high.

在i）癌症患者的治疗，ii）造血干细胞和器官移植，iii）使用 有效的免疫抑制药物和iv）先进的医疗保健已经创造了一个不断扩大的人口， 由常见环境霉菌产生的侵袭性真菌病（IFD）高危患者（曲霉病， 毛霉菌病和镰刀菌病）和正常的酵母菌病（念珠菌病）。大量研究表明 早期诊断和正确的抗真菌药物治疗将改善患者的预后。 这个转化研究项目的目标是一个多重免疫分析，将使用血清/血浆或尿液 作为诊断曲霉菌属早期感染的临床样品，镰刀菌属，毛霉目， 和念珠菌属（Candida spp.）目标患者人群将是由于以下原因而处于真菌感染高风险的个体： 使用免疫抑制药物或先进的医疗护理。该方法将是一种免疫测定法， 每种真菌的细胞壁甘露聚糖，在感染过程中流入体液。该产品将是一个 免疫分析，最有可能在侧流免疫分析（LFIA）格式，用于在初级卫生保健设置。 免疫测定将是灵敏的、特异的、快速的、廉价的和用户友好的。 有四个具体目标。目的1是进一步开发和验证高质量的单克隆抗体（mAb） 特异性针对甘露聚糖表位，所述甘露聚糖表位对于每种目标真菌群是独特的。目标2将决定 使用标本检测早期感染所需的免疫测定的分析灵敏度 从人类和小鼠模型。目标3将开发具有分析 早期诊断所需的敏感性。目的4将评估原型免疫测定的临床性能， 来自IFD动物模型和患者的血浆和尿液。 这项研究解决了一个关键的未满足的需求，诊断日益常见和致命的机会性真菌感染， 发生在高危患者身上的感染。如果成功，多重检测将成为患者的游戏规则改变者。 在乎该建议是支持一个强有力的前提，从文献和坚实的初步结果。这些 初步研究，结合现有的使用靶向脱落细胞壁的免疫测定的范例， 甘露聚糖用于诊断IFD，使得成功的概率非常高。