Detection of Virulent Burkholderia mallei

检测剧毒鼻疽伯克霍尔德菌

• 批准号: 7106509
• 负责人: STEVEN E SCHUTZER
• 金额: $ 49.43万
• 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 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7106509
• 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（调查员提供）：我们的目标是向腺体的原因Burkholderia Mallei开发区分诊断测定法。这是我们对手使用的一位经过研究的生物疗法。我们很容易受到该药物的影响，因为没有快速检测测定法，没有独特的诊断迹象和几天的短期孵育时间。另外，没有疫苗，没有感染诱导的保护性免疫，并且仅在抗生素疗效的体内数据中有限。我们认为，在使用表型型在天然宿主或人类中的表型表征的Mallei菌株鉴定出相关的核苷酸序列和抗原靶标的相关核苷酸序列和抗原靶标的相关核苷酸序列和抗原靶标，可以迅速和经济地填补。有毒或无毒的表型基于它们在宿主中杀死或产生威胁生命的疾病的能力。比较一组有毒菌株的基因组序列与无毒理（减毒）菌株可以鉴定导致这种生物毒力的独特遗传标记和区域。我们将进行平行的蛋白质组学（质谱和肽映射）研究，该研究将提供互补和收敛的数据，以查看是否表达了感兴趣的基因，并且具有生物学意义的水平。我们最初的重点是诊断。确定患者和环境中的有毒菌株以减轻威胁生命的感染并进行紧急反应非常重要。对近邻居微生物或无毒应变的反应将导致不必要且天文昂贵的响应。根据数据，我们将使用独特的核苷酸特征进行基于DNA的测试。第1阶段的原理证明将是测试设计的底漆检测和区分有毒的生物与无毒菌株和邻居附近的能力。第二个目标是在临床样品中测试这一点。在第2阶段中，我们可以执行生物学实验，以进一步开发和验证确定的诊断目标，以便在几个平台上具有实用经验，以及在第2阶段开始之前新近可用的诊断目标。在第2阶段，我们还将开发基于免疫的测定法。