Burkholderia: International Collaborative Development of Novel Diagnostics

描述（由申请人提供）： 我们将基于实时 PCR 低密度微阵列和 MLVA 开发两种 B 类病原体（鼻疽伯克霍尔德氏菌和鼻疽伯克霍尔德氏菌）的临床诊断测试，以便为医生提供快速、准确的信息。类鼻疽的病因在美国诊所很少见，但在世界许多地区（包括澳大利亚和东南亚）引起大量发病率和死亡率。鼻疽是一种人畜共患疾病，在美国也很少见。虽然这些疾病难以诊断，但早期准确地检测和识别这些疾病可以对治疗反应和疾病结果产生巨大影响。针对这些病原体开发新型诊断检测方法（实时 PCR）至关重要，因为目前临床使用的检测方法缺乏解决这些高度致命疾病所需的速度、灵敏度和特异性。系统发育分析（MLVA）将提供有关病例慢性状态以及病例之间潜在遗传联系的具体信息。 这些检测方法的开发将转化为临床和公共卫生环境，无论是单一病原体检测试剂盒还是多病原体低密度阵列格式。这些测定将提供鉴定、定量和临床重要的定性数据（例如抗生素耐药性、毒力和慢性感染标记物）。 我们的研究策略涉及多方面的转化合作，旨在优化从研究发现到临床应用的转变。这项活动的合作者包括一家非营利性研究机构 (TGen)、一所大学 (NAU)、一家临床合作伙伴（澳大利亚孟席斯/皇家达尔文医院）和一家工业/诊断制造合作伙伴（Applied Biosystems）。这一转化策略已在其他活动中证明是成功的，包括当前涉及上述三个合作伙伴（TGen、NAU 和 AB）的项目。此外，TGen 和 NAU 还与孟席斯在其他伯克霍尔德杆菌相关活动上成功合作，为该项目的无缝联盟铺平了道路。 拟议的活动将按逻辑顺序进行： A) 签名识别（具体目标#1） B) 将签名转换为测定（具体目标#2）； C) 测定验证（具体目标#3）； D) 检测制造（具体目标#4） E) 临床样品制备（具体目标#5） F) 测定的临床评估（具体目标#6） G) 分离株的高分辨率遗传分析（具体目标#7） H) 研究信息管理（具体目标#8） 我们的合作团队成员在这些领域以及这些病原体及其疾病方面都拥有丰富的经验。 简而言之，拟议的研究和开发将使用最先进的遗传技术进行新的实验室测试，以识别和描述分别引起类鼻疽和鼻疽的病原体（鼻疽伯克霍尔德氏菌和鼻疽伯克霍尔德氏菌）的感染。医学和公共卫生界将感受到开发这些新测定（测试）的重要性，因为它们将实现更快的诊断、更多有关病例的信息、更好的治疗途径以及改进疾病控制活动的流行病学信息。

DESCRIPTION (provided by applicant): We will develop clinical diagnostics tests for two Category B agents (Burkholderia pseudomallei and B. mallei) based upon real-time PCR low density microarrays and MLVA in order to provide rapid and accurate information to physicians. The etiological agent of melioidosis is rarely seen in US clinics but causes substantial morbidity and mortality in many areas of the world, including Australia and Southeast Asia. Glanders is a zoonotic disease and likewise is rarely seen in the US. While these diseases are difficult to diagnose, early and accurate detection and identification of these diseases can have a great impact on therapeutic response and disease outcomes. The development of novel diagnostic assays (real time-PCR) for these pathogens is crucial, as the current assays used in clinical settings lack the speed, sensitivity and specificity needed to address these highly fatal illnesses. The phylogenetic assay (MLVA) will provide specific information regarding the chronic status of cases, as well as the potential genetic linkage between cases. Development of these assays will translate into both clinical and public health settings, in either single pathogen assay kits or in a multiple-pathogen low density array format. These assays will provide identification, quantitative, and clinically-important qualitative data (e.g., antibiotic resistance, virulence and chronic infection markers). Our research strategy involves a multi-faceted translational collaboration, designed to optimize the move from research discovery to clinical application. The collaborators in this activity include a non-profit research institute (TGen), a university (NAU), a clinical partner (Menzies/Royal Darwin Hospital -Australia), and an industrial/diagnostics manufacturing partner (Applied Biosystems). This translational strategy has proven successful in other activities, including a current project involving three of the above partners (TGen, NAU and AB). Additionally, TGen and NAU have successfully collaborated with Menzies on other Burkholderia-re\a\ed activities, paving the way for a seamless consortium on this project. The proposed activity will move in a logical sequence: A) Signature Identification (Specific Aim #1) B) Conversion of Signatures to Assays (Specific Aim #2); C) Assay validation (Specific Aim #3); D) Assay Manufacturing (Specific Aim #4) E) Clinical Sample Preparation (Specific Aim #5) F) Clinical Evaluation of Assays (Specific Aim #6) G) High Resolution Genetic Analysis of Isolates (Specific Aim #7) H) Research Information Management (Specific Aim #8) The members of our collaborative team are highly experienced in each of these areas, as well as with these pathogens and their diseases. In short, the proposed research and development will result in novel laboratory tests, using state of the art genetic technology, for identifying and describing infections with the pathogens that cause melioidosis and glanders, Burkholderia pseudomallei and B. mallei respectively. The importance of the development of these new assays (tests) will be felt by the medical and public health community, as they will allow for faster diagnoses, more information about cases, better pathways to treatments, and improved epidemiological information for disease control activities.