Antigen Detection assay for the Diagnosis of Melioidosis

描述(申请人提供)：假腮腺伯克霍尔德氏菌是一种革兰氏阴性细菌，是类鼻疽的病原体。这种细菌在热带地区造成严重的发病率和死亡率，流行区正在扩大。类鼻疽病很难治疗和诊断，因为假鼻疽杆菌对常见的抗生素具有耐药性，而且症状是非特异性的。即使接受抗生素治疗，泰国和澳大利亚的死亡率也分别高达45%和20%。目前，还没有用于诊断类鼻疽病的有效诊断产品。患者样本培养是诊断的“金标准”；然而，这可能需要3-7天。我们的目标是开发一种快速的护理点横向流动免疫分析法(LFI)，用于直接从患者样本中检测假鼻疽囊膜抗原(CPS)。第一阶段STTR的具体目标是生产抗CPS单抗(MAbs)文库，优化LFI成分和样品制备，并对归档的类鼻疽患者样本进行可行性研究。这一第二阶段应用的目标是高级开发和验证AMD LFI原型，该原型结合了更高亲和力的CPS单抗。通过完成这些目标，我们希望生产出一种优化的LFI，其灵敏度等于或高于患者样本的培养。为了实现这些目标，我们建立了一支经验丰富的团队。Aucoin博士的实验室和InBios International之间存在着强有力的合作伙伴关系，这导致了令人鼓舞的LFI原型的开发。此外，两个小组将与流行地区(泰国和澳大利亚)的专家密切合作，后者将对不同患者样本类型的LFI进行大规模临床前评估。

DESCRIPTION (provided by applicant): Burkholderia pseudomallei is a Gram-negative bacterium that is the causative agent of melioidosis. The bacterium causes significant morbidity and mortality in tropical regions and endemic areas are expanding. Melioidosis is difficult to trea and diagnose due to the fact that B. pseudomallei is resistant to common antibiotics and symptoms are non-specific. Mortality rates reach 45% and 20% in Thailand and Australia respectively, even with antibiotic treatment. Currently, there is no validated diagnostic product for the diagnosis of melioidosis. Culturing of patient samples is the "gold standard" for diagnosis; however this can take 3-7 days. Our goal is to develop a rapid point-of-care lateral flow immunoassay (LFI) for the detection of the B. pseudomallei capsular antigen (CPS) directly from patient samples. Phase I STTR Specific Aims focused on producing a library of anti-CPS monoclonal antibodies (mAbs), optimizing the LFI components and sample preparation, and performing a feasibility study on archived melioidosis patient samples. The objectives of this Phase II application is advanced development and validation of the prototype AMD LFI that incorporates a higher affinity CPS mAb. By completing these goals we hope to produce an optimized LFI that possesses sensitivity equal to or greater than culturing of patient samples. To achieve these goals we have established an experienced team. A strong partnership exists between Dr. AuCoin's laboratory and InBios International that has led to the development of an encouraging prototype LFI. In addition, both groups will work closely with experts in endemic areas (Thailand and Australia) who will be conducting a large preclinical evaluation of the LFI with different patient sample types.