Detection of Virulent Burkholderia mallei

描述(由研究人员提供)：我们的目标是开发对马来伯克霍尔德氏菌的鉴别诊断分析，马来伯克霍尔德氏菌是扁桃体的原因。这是一种被我们的对手使用的未被充分研究的生物制剂。我们很容易受到这种病原体的影响，因为没有快速检测，没有明显的诊断迹象，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，在确定相关的核苷酸序列和抗原靶标后，使用自然宿主马或人的表型特征的马利氏杆菌菌株，采用完整的比较基因组和蛋白质组方法的新组合，可以迅速和经济地填补这些空白。强毒或无毒的表型取决于它们在宿主中杀死或产生威胁生命的疾病的能力。比较一组毒力菌株和弱毒(弱毒)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(质谱学和肽图谱)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，以及是否在生物意义的水平上。我们最初的重点是诊断学。在患者和环境中确定一种毒力菌株非常重要，以减轻危及生命的感染并采取紧急反应。对近邻微生物或无毒菌株的反应将导致不必要的反应和天文数字般的昂贵。根据这些数据，我们将使用独特的核苷酸签名进行基于DNA的测试。第一阶段的原则证明将是测试设计的引物检测和区分有毒有机物与无毒菌株和近邻的能力。第二个目标将是在临床样本中测试这一点。在第二阶段，我们可以进行生物实验，在我们有实际经验的几个平台上进一步开发和验证已识别的诊断目标，以及那些可能在第二阶段开始之前新推出的平台。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.