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“黄金标准”，但这可能需要3-5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方法，用于直接从患者样本中检测假鼻疽囊膜抗原(CPS)。我们之前的研究已经确定CPS是一种脱落抗原，LFI的原型已经产生了令人鼓舞的结果。该项目的目标是(I)建立高亲和力CPS特异性单抗文库，(Ii)优化LFI结构，(Iii)确定相容的样本类型和样本制备，以及(Iv)定量患者样本中的CPS。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博士的实验室已经在与InBios合作开发LFI原型。此外，这两个小组将与流行地区(泰国和澳大利亚)的专家密切合作，他们将用不同的样本类型对LFI进行评估。 公共卫生相关性：类鼻疽病是一种进展迅速的疾病，如果不治疗，往往是致命的。假鼻疽伯克霍尔德氏菌是类鼻疽的病原体，这种土壤寄生细菌已被列为B类选择剂。在流行地区(泰国、东南亚和澳大利亚北部)，类鼻疽病是一个公共卫生问题。需要对类鼻疽病进行快速诊断，以便适当地使用抗生素进行有效治疗。目前，诊断的“金标准”是患者样本的培养，这需要3-5天的时间，并大大推迟了适当治疗的实施。在这项应用中，我们提出开发一种针对类鼻疽病的具有成本效益的快速诊断方法，可以在几分钟内得出结果。这种快速诊断测试可以帮助检测和监测现场的急性和慢性假鼻疽杆菌感染，无论是在流行地区还是在生物恐怖袭击之后。

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 3-5 days. Our goal is to develop a rapid point-of-care lateral flow immunoassay for the detection of the B. pseudomallei capsular antigen (CPS) directly from patient samples. Our previous studies have determined that the CPS is a shed antigen and a prototype LFI has yielded encouraging results. The goals of the project are to (i) produce a library of high affinity CPS specific mAbs (ii) optimize LFI construction, (iii) determine compatibe sample types and sample preparation and (iv) quantify CPS in patient samples. 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. Dr. AuCoin's laboratory is already collaborating with InBios on development of a prototype LFI. In addition, both groups will work closely with experts in endemic areas (Thailand and Australia) who will be conducting evaluation of the LFI with different sample types. PUBLIC HEALTH RELEVANCE: Melioidosis is a rapidly progressing and frequently fatal disease if left untreated. Burkholderia pseudomallei is the causative agent of melioidosis, this soil dwelling bacterium has been listed as a Category B Select Agent. In the endemic areas (Thailand, Southeast Asia and northern parts of Australia), melioidosis is a public health concern. Rapid diagnosis of melioidosis is needed in order to properly administer effective treatment with antibiotics. At present, the diagnostic "gold standard" is culturing of patient samples, which takes 3-5 days and significantly delays administration of proper treatment. In this application, we offer to develop a point-of-care, cost-effective, and rapid diagnostic for melioidosis that can deliver results within minutes. Such rapid diagnostic test can assist in detecting and monitoring acute and chronic B. pseudomallei infections in the field both in endemic areas as well as following a bioterrorist attack.