Burkholderia pseudomallei antimicrobial resistance mechanisms

因为B。由于类鼻疽（Bp's）对许多抗生素具有内在耐药性，类鼻疽病的治疗很困难 并且必须持续延长的时间段。有限的抗生素谱可用于 类鼻疽的治疗，以及抗生素治疗期间耐药菌株的出现，需要更好的 了解潜在的耐药机制，以便能够进行适当的治疗干预。 此外，成功和适当地治疗由涉及菌株的生物恐怖事件引起的感染， 通过自然手段或恶意基因工程获得抗性决定因素， 如果不能容易地识别潜在的耐药机制，则可能不可能。这个假设是 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 启动或重新定向（在类鼻疽治疗期间的临床环境中）。将追求三个目标，以测试 假设： 目的1 -确定临床和环境分离株的耐药机制。RT-PCR，表达 克隆的基因产物和生物化学方法将用于表征抗性决定因素 来自对β-内酰胺类、多西环素、甲氧苄啶和 磺胺甲恶唑目的2 -基因工程菌株耐药机制的定义。这将 通过表征转座子诱导的抗性突变体，以及通过选择和 表征对AIM 1中测试的抗生素的自发抗性突变体。目标3 -设计 定向DNA微阵列作为快速鉴定抗性决定簇的工具。利用信息 从目标1和2中收集，将产生定向DNA微阵列，其与一组 特异性PCR引物，将提供一套快速确定抗性机制的诊断工具 在不同来源的Bp分离物中。 BP是RMRCE细菌治疗综合研究重点的一个组成部分，其主要工作是 是为了鉴定这种病原体和其他一些选择剂的新疗法。的 本研究中获得的工具、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.