Burkholderia: International Collaborative Development of Novel Diagnostics

描述(由申请人提供)： 我们将开发基于实时聚合酶链式反应低密度芯片和MLVA的两种B类病原体(假鼻疽伯克霍尔德氏菌和马来伯克霍尔德菌)的临床诊断测试，以便为医生提供快速和准确的信息。类鼻疽病的病原体在美国的临床上很少见，但在世界许多地区，包括澳大利亚和东南亚，会导致大量的发病率和死亡率。扁桃体是一种人畜共患疾病，同样在美国也很少见。虽然这些疾病很难诊断，但早期和准确地发现和识别这些疾病可以对治疗反应和疾病结局产生重大影响。开发针对这些病原体的新型诊断分析方法(实时聚合酶链式反应)至关重要，因为目前临床上使用的分析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的具体信息，以及病例之间潜在的遗传联系。 这些检测的开发将转化为临床和公共卫生环境，无论是单一病原体分析试剂盒还是多病原体低密度阵列格式。这些检测将提供鉴定、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过程。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)、一个临床合作伙伴(Menzies/澳大利亚皇家达尔文医院)和一个工业/诊断制造合作伙伴(应用生物系统)。这一翻译战略在其他活动中已被证明是成功的，包括目前涉及上述三个伙伴(TGen、NAU和AB)的一个项目。此外，TGen和NAU已经成功地与Menzies在Burkholderia Re\a\a的其他活动上进行了合作，为这个项目的无缝联盟铺平了道路。 建议的活动将按逻辑顺序进行： A)签名识别(具体目标1) B)将签名转换为化验(具体目标2)； C)分析验证(具体目标3)； D)分析制造(具体目标4) E)临床样本制备(具体目标5) F)化验的临床评价(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都非常有经验。 简而言之，拟议的研究和开发将导致使用最先进的基因技术进行新的实验室测试，以识别和描述分别导致类鼻疽病和腺体的病原体--假鼻疽伯克霍尔德氏菌和马来伯克霍尔德氏菌的感染情况。医学界和公共卫生界将感受到开发这些新的分析(测试)的重要性，因为它们将允许更快地诊断、更多关于病例的信息、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.