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: 10269054
• 负责人: Thomas R Kozel
• 金额: $ 68.15万
• 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/10269054
• 关键词: Address; Advanced Development; Animal Model; Antibodies; Antifungal Agents; Archives; Aspergillosis; Aspergillus; 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; Immunosuppressive Agents; 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; Plant Roots; Plasma; Polysaccharides; Primary Health Care; Probability; Production; Rapid diagnostics; 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; biological specimen archives; cancer therapy; fungus; galactomannan; health care settings; high risk; human model; improved; monoclonal antibody production; mortality; mouse model; multiplex assay; 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.

1)癌症患者治疗的最新进展，2)造血干细胞和器官移植，3)应用 强大的免疫抑制药物的使用和先进的医疗保健创造了不断增长的人口 由常见环境霉菌(曲霉病、 毛霉病和丝孢菌病)和正常共生酵母(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。 这一转化研究项目的目标是一种使用血清/血浆或尿液的多重免疫分析。 作为诊断曲霉属、镰刀菌属、毛霉属早期感染的临床标本 和假丝酵母属(Candida spp.)目标患者人群将是因以下原因而感染真菌的高危人群 使用免疫抑制药物或高级医疗护理。这种方法将是一种免疫分析，可以检测到 每种真菌的细胞壁甘露糖在感染期间被释放到体液中。该产品将是一个 免疫分析，最有可能是横向流动免疫分析(LFIA)格式，用于初级卫生保健环境。 免疫分析将是灵敏、特异、快速、廉价和用户友好的。 有四个具体目标。AIM 1将进一步开发和验证高质量的单抗(MAbs) 对于每个目标真菌组所特有的甘露聚糖表位。目标2将决定 利用标本检测早期感染所需的免疫分析的灵敏度 来自人类和小鼠模型。目标3将开发具有分析能力的多重免疫分析方法 早期诊断所需的敏感性。AIM 4将评估原型免疫分析的临床性能，使用 动物模型和IFD患者的血浆和尿液。 这项研究解决了诊断日益常见和致命的机会性真菌的一个关键的未得到满足的需求 发生在高危患者身上的感染。如果成功，多路化验将改变患者的游戏规则 关心。这一建议得到了来自文献的强有力的前提和坚实的初步结果的支持。这些 初步研究，结合使用针对脱落细胞壁的免疫分析的现有范例 甘露醇对IFD的诊断，使成功的概率很高。