Burkholderia pseudomallei antimicrobial resistance mechanisms

由于假鼻疽杆菌(BP)对多种抗生素的固有耐药性，类鼻疽病的治疗很困难。 并且必须持续很长一段时间。可用于治疗的抗生素的有限光谱 类鼻疽的治疗，以及在抗生素治疗过程中出现耐药菌株，需要更好的 了解潜在的耐药性机制，以便进行适当的治疗干预。 此外，成功和适当地治疗涉及菌株的生物恐怖主义事件造成的感染 获得了抗性决定因素，无论是通过自然手段还是通过恶意基因工程， 如果不能很容易地确定潜在的抵抗机制(S)，可能是不可能的。假设是 对一线临床药物耐药机制的定义将允许设计 快速检测临床分离株或恶意工程菌耐药机制的策略 菌株。反过来，可以及早发现耐药性(生物恐怖主义事件)并进行适当的治疗 启动或重定向(在类鼻疽病治疗期间的临床环境中)。将追求三个目标来测试 假设： 目的1-确定临床分离株和环境分离株的耐药机制。逆转录聚合酶链式反应，表达 克隆的基因产物和生化方法将被用来表征抗性决定因素。 从临床和环境中分离出对β-内酰胺类、多西环素、甲氧苄氨嘧啶和 磺胺甲恶唑。目的2-定义基因工程菌株的抗药性机制。这将是 通过鉴定转座子诱导的抗性突变体，以及通过选择和 对目标1中测试的抗生素的自发耐药突变体的特征。目标3-设计一种 将DNA微阵列作为一种快速鉴定抗性决定因素的工具。利用信息 从AIMS 1和2收集的定向DNA微阵列将与一组 特异的聚合酶链式反应，为快速确定抗性机制提供了一套诊断工具 来自不同来源的BP中的分离株。 BP是RMRCE细菌治疗综合研究重点的组成部分，其主要努力 旨在鉴定针对这种病原体和其他一些选择药物的新疗法。这个 在这项研究中获得的工具、突变体和知识将直接有益于这些努力。

Because of B. pseudomallefs (Bp's) intrinsic resistance to many antibiotics, melioidosis therapy is difficult and must be continued for extended periods of time. The limited spectrum of antibiotics available for melioidosis treatment, and the emergence of resistant strains during antibiotic therapy call for a better understanding of underlying resistance mechanisms to enable proper therapeutic interventions. Furthermore, successful and proper treatment of infections caused by bioterrorism events involving strains having acquired resistance determinants, whether by natural means or by malicious genetic engineering, may be impossible if the underlying resistance mechanism(s) cannot be readily identified. The hypothesis is that a definition of the mechanisms governing resistance to frontline clinical drugs will allow design of strategies for rapid detection of resistance mechanisms in clinical isolates or in maliciously engineered strains. In turn, then, resistance can be detected early (bioterrorism events) and appropriate treatment initiated or redirected (in clinical settings during melioidosis therapy). Three aims will be pursued to test the hypothesis: Aim 1 - Definition of resistance mechanisms in clinical and environmental isolates. RT-PCR, expression of cloned gene products and biochemical approaches will be employed to characterize resistance determinants from clinical and environmental isolates resistant to p-lactams, doxycyline, trimethoprim and sulfamethoxazole. Aim 2 - Definition of resistance mechanisms in genetically engineered strains. This will be achieved by characterizing transposon-induced resistant mutants, as well as by selecting and characterizing spontaneously resistant mutants to the antibiotics tested in aim 1. Aim 3 - Design of a directed DNA microarrav as a tool for rapid identification of resistance determinants. Utilizing the information gleaned from aims 1 and 2, a directed DNA microarray will be generated which, in concert with a set of specific PCR primers, will provide a set of diagnostic tools for rapid determination of resistance mechanism in Bp isolates from diverse sources. Bp is an integral part of the RMRCE Bacterial Therapeutics Integrated Research Focus , whose main efforts are geared towards identification of novel therapeutics for this pathogen and a few other Select Agents. The tools, mutants and knowledged gained in this study will directly benefit these efforts.