Detection of Virulent Burkholderia mallei

检测剧毒鼻疽伯克霍尔德菌

• 批准号: 6992272
• 负责人: STEVEN E SCHUTZER
• 金额: $ 49.42万
• 依托单位: BIOSCIENCE DEVELOPMENT, INC.
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-15 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6992272
• 关键词: Burkholderia; DNA primers; bacterial DNA; bacterial genetics; biohazard detection; bioterrorism /chemical warfare; gene expression; genetic strain; mass spectrometry; proteomics; technology /technique development; virulence

DESCRIPTION (provided by investigator): Our objective is to develop discriminating diagnostic assays to Burkholderia mallei, the cause of glanders. It is an understudied biothreat agent that has been used by our adversaries. We are vulnerable to this agent because there is no rapid detection assay, no distinctive diagnostic signs, and a short incubation period of days. In addition there is no vaccine, no infection-induced protective immunity, and only limited reliable in vivo data on antibiotic efficacy. We believe these gaps can be filled expeditiously and economically after identifying relevant nucleotide sequence and antigenic targets with a novel combination of complete comparative genomic and proteomic approach using phenotypically characterized B. mallei strains in the natural host- equine or human. The phenotype of virulent or avirulent is based on their ability to kill or produce life threatening disease in the host. Comparison of the genomic sequences of a set of virulent strains to avirulent (attenuated) strains can identify unique genetic markers and regions responsible for the virulence of this organism. We will perform parallel proteomic (mass spectrometry and peptide mapping) studies which will provide complementary and convergent data to see if the gene of interest is being expressed and at a level of biologic significance. Our initial focus is on diagnostics. It is very important to identify a virulent strain in the patient and the environment to mitigate against life-threatening infections and to mount an emergency response. A response to a near neighbor microbe or an avirulent strain will result in an unnecessary and astronomically expensive response. Based on the data, we will use unique nucleotide signatures for a DNA based test. Proof of principle for Phase 1 will be to test the ability of designed primers to detect and discriminate the virulent organism from the avirulent strain and near neighbors. A second goal will be to test this in clinical samples. In Phase 2 we can perform biologic experiments to further develop and validate identified targets for diagnostics in several platforms with which we have practical experience and those which may be newly available before Phase 2 begins. In Phase 2 we will also develop the immuno-based assays.

描述（由研究者提供）：我们的目标是针对鼻疽病的病因鼻疽伯克霍尔德氏菌开发鉴别诊断方法。它是一种未经充分研究的生物威胁剂，已被我们的对手使用。我们很容易受到这种物质的影响，因为没有快速检测方法，没有明显的诊断标志，而且潜伏期很短。此外，没有疫苗，没有感染引起的保护性免疫，关于抗生素功效的可靠体内数据也有限。我们相信，在使用天然宿主马或人中表型特征的鼻疽杆菌菌株，通过完整的比较基因组和蛋白质组学方法的新组合鉴定相关核苷酸序列和抗原靶标后，可以迅速且经济地填补这些空白。有毒或无毒的表型取决于它们杀死宿主或在宿主中产生危及生命的疾病的能力。比较一组强毒菌株与无毒（减毒）菌株的基因组序列可以识别独特的遗传标记和负责该生物体毒力的区域。我们将进行平行的蛋白质组学（质谱和肽图）研究，这将提供互补和收敛的数据，以查看感兴趣的基因是否正在表达以及是否处于生物学意义的水平。我们最初的重点是诊断。识别患者和环境中的毒力菌株对于减轻危及生命的感染并采取紧急应对措施非常重要。对邻近微生物或无毒菌株的反应将导致不必要且昂贵的反应。根据这些数据，我们将使用独特的核苷酸特征进行基于 DNA 的测试。第一阶段的原理验证将测试设计的引物检测和区分有毒微生物与无毒菌株和邻近菌株的能力。第二个目标是在临床样本中进行测试。在第二阶段，我们可以进行生物实验，以在我们拥有实践经验的几个平台以及第二阶段开始之前可能新推出的平台上进一步开发和验证已确定的诊断目标。在第二阶段，我们还将开发基于免疫的检测方法。