Immunoassay for capsular antigen for rapid diagnosis of anthrax

说明(申请人提供)：只有在感染的非常早期阶段使用抗生素才能有效对抗炭疽病，此时症状非特异性，诊断困难。因此，早期诊断是治疗新生儿感染的医学对策的核心。 炭疽杆菌被一个由聚-D-谷氨酸(PGA)组成的胶囊所包围。初步研究发现，在肺炭疽病动物模型中，PGA被释放到血清和尿液中，PGA免疫分析是血培养诊断的有效替代方法。PGA出现在血液样本将产生阳性血培养的时间或之前。然而，与血液培养可能需要几天时间才能产生阳性结果不同，快速格式的免疫分析可以在几秒钟到几分钟内产生阳性结果。更多的研究发现，EIA可以成功地迁移到侧向流动免疫层析(LFI)格式。LFI检测平台具有快速、高灵敏度、易于使用、适应性强和成本效益高的特点。 总体假设是，血和尿中PGA的免疫测定是诊断炭疽病血培养的有效替代方法。总体目标是将该项目从概念验证转移到LFI格式的免疫分析的优化，并使用人体样本和兔子和食蟹猴的吸入性炭疽模型对该分析进行验证。其具体目标是：i)生产和优化LFI格式的免疫分析；ii)在实验室环境中评估和验证免疫分析性能；iii)使用动物模型和人类炭疽病样本验证免疫分析性能。 这是一个成熟的项目，已经从i)潜在诊断靶点的最初概念到ii)研究级免疫分析的发展到iii)动物模型中的概念验证靶标验证到iv)以护理点形式生产原型免疫分析。如果成功，拟议的诊断工具将大大加强对生物恐怖主义事件的准备。

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.