Burkholderia pseudomallei antimicrobial resistance mechanisms

类鼻疽伯克霍尔德氏菌耐药机制

• 批准号: 8261423
• 负责人: HERBERT P. SCHWEIZER
• 金额: $ 24.48万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8261423
• 关键词: Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antimicrobial Resistance; Bacteria; Biochemical; Bioterrorism; Burkholderia pseudomallei; Clinical; DNA; DNA Microarray Chip; Diagnostic; Engineering; Event; Genetic Engineering; Glean; Infection; Lactams; Learning; Melioidosis; Pharmaceutical Preparations; Proteins; Research; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Site; Source; Testing; Therapeutic; Therapeutic Intervention; Time; Trimethoprim-Sulfamethoxazole; biodefense; design; drug discovery; expression cloning; mutant; novel therapeutics; pathogen; rapid detection; resistance mechanism; resistant strain; tool

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.

由于假杆菌（BP）对许多抗生素的固有抗药性，因此很难 并且必须长时间继续。有限的抗生素光谱可用于 Melioidosis治疗以及抗生素治疗期间抗性菌株的出现要求更好 了解潜在的抗性机制，以实现适当的治疗干预措施。 此外，由涉及菌株的生物恐怖事件引起的成功和适当治疗感染 获得了抵抗决定因素，无论是自然手段还是通过恶意基因工程， 如果无法轻易确定潜在的抵抗机制，则可能是不可能的。该假设是 关于对一线临床药物的抵抗力的机制的定义将允许设计 快速检测临床分离株或恶意工程中的耐药机制的策略 菌株。然后，可以尽早检测到抵抗力（生物恐怖事件）和适当的治疗 启动或重定向（在黑梅利病治疗期间的临床环境中）。将追求三个目标来测试 假设： 目标1-临床和环境分离株中抗性机制的定义。 RT-PCR，表达 克隆的基因产物和生化方法将用于表征抗药性决定因素 从抗p-内酰胺，强力线，甲氧苄啶和 亚甲氧唑。 AIM 2-基因工程菌株中抗性机制的定义。这会 可以通过表征转座子诱导的抗性突变体以及选择和选择和选择 表征在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 Microarrav是快速识别抗性决定因素的工具。利用信息 将从目标1和2中收集，将产生一个定向的DNA微阵列，并与一组一组 特定的PCR引物将提供一组诊断工具，用于快速确定电阻机制 在来自不同来源的BP分离株中。 BP是RMRCE细菌疗法综合研究重点的组成部分，其主要努力 旨在鉴定该病原体和其他一些精选药物的新型治疗剂。这 在这项研究中获得的工具，突变体和知识者将直接受益于这些努力。