Detection of Virulent Burkholderia mallei

描述（由研究者提供）：我们的目标是开发对引起腺体病变的mallei伯克霍尔德氏菌的鉴别诊断方法。这是一种未经充分研究的生物威胁剂我们的对手已经使用过了。我们很容易受到这种药物的影响，因为它没有快速检测方法，没有明显的诊断体征，而且潜伏期很短。此外，没有疫苗，没有感染诱导的保护性免疫，只有有限的可靠的体内抗生素疗效数据。我们相信，在利用自然宿主马或人类中具有表型特征的马氏B. mallei菌株的完整比较基因组学和蛋白质组学方法的新组合确定相关的核苷酸序列和抗原靶点后，这些空白可以迅速而经济地填补。毒性或无毒性的表型是基于它们在宿主中杀死或产生危及生命的疾病的能力。将一组毒株与无毒（减毒）毒株的基因组序列进行比较，可以确定与该生物毒力有关的独特遗传标记和区域。我们将进行平行蛋白质组学（质谱和肽图谱）研究，这将提供互补和趋同的数据，以了解感兴趣的基因是否正在表达，以及是否具有生物学意义。我们最初的重点是诊断。识别患者和环境中的毒性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。对附近的微生物或无毒菌株的反应将导致不必要的和天文数字昂贵的反应。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。第一阶段的原理证明将是测试设计的引物检测和区分毒性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，以进一步开发和验证我们有实践经验的几个平台上确定的诊断目标，以及在第二阶段开始之前可能新提供的平台。在第二阶段，我们还将开发基于免疫的检测方法。

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.