Burkholderia: International Collaborative Development of Novel Diagnostics

描述（由申请人提供）： 我们将开发两种B类病原体（类鼻疽伯克霍尔德菌和B类）的临床诊断检测。mallei）的实时PCR低密度微阵列和MLVA的基础上，以便提供快速和准确的信息给医生。类鼻疽的病原体在美国临床上很少见，但在世界许多地区（包括澳大利亚和东南亚）引起大量的发病率和死亡率。腺是一种人畜共患疾病，在美国也很少见。虽然这些疾病很难诊断，但这些疾病的早期和准确检测和识别可以对治疗反应和疾病结果产生重大影响。开发针对这些病原体的新型诊断检测方法（真实的时间PCR）至关重要，因为目前用于临床环境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或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、毒力和慢性感染标志物）。 我们的研究战略涉及多方面的翻译合作，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个非营利性研究机构（TGen）、一所大学（NAU）、一个临床合作伙伴（Menzies/皇家达尔文医院-澳大利亚）和一个工业/诊断制造合作伙伴（应用生物系统公司）。这一转化战略已在其他活动中证明是成功的，包括目前涉及上述三个合作伙伴（TGen、NAU和AB）的项目。此外，TGen和NAU还成功地与孟席斯在其他伯克霍尔德氏菌相关活动上进行了合作，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）测定生产（具体目标#4） E）临床样品制备（具体目标#5） F）试验的临床评价（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） 我们合作团队的成员在这些领域以及这些病原体及其疾病方面经验丰富。 简而言之，拟议的研究和开发将导致新的实验室检测，使用最先进的遗传技术，用于识别和描述引起类鼻疽和鼻疽的病原体，类鼻疽伯克霍尔德氏菌和B的感染。鼻疽分别。医学和公共卫生界将感受到开发这些新检测方法的重要性，因为它们将使诊断速度更快，病例信息更多，治疗途径更好，并为疾病控制活动提供更好的流行病学信息。

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.