Immunoassay for capsular antigen for rapid diagnosis of anthrax

荚膜抗原免疫分析快速诊断炭疽病

• 批准号: 8076076
• 负责人: Thomas R Kozel
• 金额: $ 71.81万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8076076
• 关键词: Advanced Development; Aerosols; Animal Model; Animals; Anthrax disease; Antibiotics; Antigens; Bacillus anthracis; Bacteria; Biological Assay; Bioterrorism; Blood; Blood specimen; Breathing; Detection; Development; Diagnosis; Diagnostic; Diagnostic tests; Early Diagnosis; Glutamic Acid; Goals; Human; Immunoassay; Infection; Laboratories; Lateral; Lung; Macaca; Medical; Modeling; Oryctolagus cuniculus; Patients; Performance; Plasma; Production; Readiness; Reproducibility; Research; Sampling; Serum; Specificity; Specimen; Staging; Symptoms; Testing; Time; Urine; Validation; base; capsule; cost effective; design; nonhuman primate; point of care; prototype; rapid diagnosis; response; tool

DESCRIPTION (provided by applicant): Antibiotics are effective against anthrax only if given at a very early stage of infection when symptoms are non-specific and diagnosis is difficult. As a consequence, early diagnosis is central to medical countermeasures designed to treat treatment nascent infection. Bacillus anthracis is surrounded by a capsule composed of poly-?-D-glutamic acid (PGA). Preliminary studies found that PGA is shed into serum and urine during animal models of pulmonary anthrax and that immunoassay for PGA is an effective surrogate for the blood culture in diagnosis. PGA appears in blood at or before the time that a blood sample will yield a positive blood culture. However, unlike the blood culture which can take several days to produce positive results, immunoassay in rapid format can produce positive results in seconds to minutes. Additional studies found that the EIA can be successfully migrated to a lateral flow immunochromatographic (LFI) format. The LFI assay platform is rapid, highly sensitive, easy to use, highly adaptable, and cost effective. The overall hypothesis is that immunoassay for PGA in blood and urine is an effective surrogate for the blood culture in diagnosis of anthrax. The overall goal is to move the project from proof-of-concept to optimization of an immunoassay in LFI format and validation of the assay using human samples and models of inhalation anthrax in rabbits and Cynomolgus macaques. The Specific Aims are to i) produce and optimize an immunoassay in LFI format, ii) evaluate and validate immunoassay performance in a laboratory setting and iii) validate immunoassay performance using animal models and samples from human anthrax. This is a mature project that has moved from i) an initial concept for a potential diagnostic target to ii) development a of research-grade immunoassay to iii) proof-of-concept target validation in animal models to iv) production of a prototype immunoassay in point-of-care format. If successful, the proposed diagnostic tool will significantly enhance readiness for a bioterrorism incident.

描述（由申请人提供）：只有在感染的早期阶段使用抗生素才能有效对抗炭疽，此时症状不具特异性且诊断困难。因此，早期诊断对于旨在治疗新生感染的医疗对策至关重要。 炭疽杆菌被由聚-β-D-谷氨酸（PGA）组成的胶囊包围。初步研究发现，在肺炭疽动物模型中，PGA 会进入血清和尿液中，PGA 免疫检测是血培养诊断的有效替代方法。 PGA 在血样产生阳性血培养物时或之前出现在血液中。然而，与血培养需要几天才能产生阳性结果不同，快速免疫测定可以在几秒到几分钟内产生阳性结果。其他研究发现 EIA 可以成功迁移到侧流免疫色谱 (LFI) 格式。 LFI 检测平台快速、高灵敏度、易于使用、适应性强且具有成本效益。 总体假设是，血液和尿液中 PGA 的免疫测定是诊断炭疽病时血培养的有效替代方法。总体目标是将项目从概念验证转向优化 LFI 格式的免疫测定，并使用人体样本以及兔子和食蟹猴吸入炭疽模型验证测定。具体目标是 i) 以 LFI 格式生成和优化免疫测定，ii) 在实验室环境中评估和验证免疫测定性能，以及 iii) 使用动物模型和人类炭疽样本验证免疫测定性能。 这是一个成熟的项目，从 i) 潜在诊断靶标的初步概念，到 ii) 研究级免疫测定的开发，到 iii) 动物模型中的概念验证靶标验证，再到 iv) 原型免疫测定的生产采用即时护理格式。如果成功，拟议的诊断工具将显着增强对生物恐怖主义事件的准备。