Beta-lactamase fluorescent probes for bacterial detection

用于细菌检测的 β-内酰胺酶荧光探针

• 批准号: 9309417
• 负责人: Jianghong Rao
• 金额: $ 58.42万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-01-10 至 2020-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9309417
• 关键词: Acid Fast Bacillae Staining Method; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Antimicrobial Resistance; Bacillus (bacterium); Bacteria; Bacterial Infections; Biological; Biological Assay; Biological Markers; Carbapenems; Cephalosporins; Cessation of life; Chemistry; Clinical; Clinical Microbiology; Complex; Computer Simulation; Dangerousness; Detection; Development; Diagnostic Procedure; Diagnostic tests; Disease; Drug resistance; Enterobacteriaceae; Enzymes; Escherichia coli; Fluorescence; Fluorescent Probes; Goals; Growth; Health Care Costs; Health Professional; Human; Hydrolase; In Vitro; Individual; Infection; Klebsiella pneumonia bacterium; Lactams; Left; Methods; Molecular Probes; Monobactams; Mycobacterium tuberculosis; Organism; Outcomes Research; Pathogenicity; Patients; Population; Prevention; Prevention strategy; Production; Public Health; Recording of previous events; Reporting; Research; Research Personnel; Resistance; Resources; Sampling; Series; Signal Transduction; Specificity; Sputum; Testing; Time; Tuberculosis; United States; Virus Diseases; Work; antimicrobial drug; assay development; beta-Lactam Resistance; beta-Lactamase; beta-Lactams; carbapenem-resistant Enterobacteriaceae; carbapenemase; chemical synthesis; cost; cost effective; design; effective therapy; healthcare community; improved; killings; member; novel; pathogen; pathogenic bacteria; pressure; prevent; rapid detection; rapid technique; resistant strain; respiratory; stereochemistry; transmission process; treatment strategy

Project Summary In September 2014, President Barack Obama signed an executive order directing federal resources to improve detection and prevention of antibiotic resistance, with a goal of developing a diagnostic test by 2020 that can distinguish between bacterial and viral infections in at least 20 min. The long-term goal of this research is to provide a novel platform solution for rapid, sensitive and specific detection of pathogenic bacterial infections through development of fluorescent probes specific to individual β-lactamases expressed in the bacteria. β-Lactamases are a class of bacterial hydrolases destroying β-lactam antibiotics and rendering antibiotics resistance in bacterial pathogens. Today many patients with suspected infections are administered antibiotics (most frequently β-lactams) empirically without prior proper identification of the causative agent, resulting in antibiotics overuse and overspread of antimicrobial resistance, with >500,000 deaths in the world annually attributable to these infections. This proposal will focus on two deadly bacteria: Mycobacterium tuberculosis (Mtb) and carbapenem-resistant Enterobacteriaceae (CRE). Tuberculosis represents one of the most dangerous respiratory pathogens in the history of mankind, killing over one million people each year and infecting one third of the world's population. Tubercle bacilli express BlaC, an Ambler class A β-lactamase, so Mtb has the intrinsic resistance to β-lactam antibiotics. Accumulating results including those from us have supported the use of BlaC as the biomarker for Mtb detection. Broad-spectrum carbapenem agents are frequently the last option for effective therapy of infections with antimicrobial resistant organisms, but the emergence of carbapenem-resistant Enterobacteriaceae (CRE) over the past decade has left clinicians with few treatment options. CRE is frequently due to the production of carbapenemase enzymes that efficiently hydrolyze carbapenems and other β-lactam antibiotics. This project will explore novel double-quenching, dual targeting probe design strategy to develop BlaC-specific substrate probes for rapid, highly sensitive Mtb detection (Aim #1) and carbapenemase-specific substrate probes for rapid, highly sensitive CRE detection (Aim #2). These new probes will be evaluated with clinical patient samples. The expected outcome of this research is that a series of novel fluorescent probes will be discovered as a novel assay platform to enable rapid, sensitive and specific detection of Mtb and CRE. We envision that these assays could reduce unnecessary healthcare costs associated with treatment of bacterial infection and prevent further expansion of existing drug-resistant strains.

项目概要 2014 年 9 月，巴拉克·奥巴马总统签署了一项行政命令，指示联邦资源改善 检测和预防抗生素耐药性，目标是到 2020 年开发出一种诊断测试，可以 至少 20 分钟内区分细菌和病毒感染 这项研究的长期目标是 为快速、灵敏、特异的病原菌感染检测提供新颖的平台解决方案 通过开发针对细菌中表达的单个β-内酰胺酶的特异性荧光探针。 β-内酰胺酶是一类细菌水解酶，可破坏β-内酰胺抗生素并产生抗生素 如今，许多疑似感染的患者都接受了抗生素治疗。 （最常见的是β-内酰胺）凭经验而没有事先正确识别病原体，导致 抗生素的过度使用和抗菌素耐药性的蔓延，每年导致全球超过 50 万人死亡 归因于这些感染。 该提案将重点关注两种致命细菌：结核分枝杆菌 (Mtb) 和碳青霉烯类抗药性细菌 肠杆菌科 (CRE) 是最危险的呼吸道病原体之一。 人类历史上每年造成超过一百万人死亡并感染世界三分之一的人口。 结核杆菌表达BlaC，一种Ambler A类β-内酰胺酶，因此Mtb对β-内酰胺具有内在耐药性 包括我们在内的累积结果支持使用 BlaC 作为抗生素的生物标志物。 结核分枝杆菌检测。 广谱碳青霉烯类药物通常是有效治疗感染的最后选择 抗菌药物耐药微生物，但碳青霉烯类耐药肠杆菌科细菌 (CRE) 的出现超过 过去十年，CRE 的治疗选择很少，这通常是由于生产的原因。 碳青霉烯酶可有效水解碳青霉烯类和其他 β-内酰胺抗生素。 该项目将探索新颖的双淬灭、双靶向探针设计策略，以开发 BlaC 特异性 用于快速、高灵敏度 Mtb 检测（目标 #1）和碳青霉烯酶特异性底物的底物探针 用于快速、高灵敏度 CRE 检测的探针（目标#2）将通过临床评估这些新探针。 患者样本。 这项研究的预期结果是，将发现一系列新型荧光探针作为 新型检测平台可实现 Mtb 和 CRE 的快速、灵敏和特异性检测。 检测可以减少与治疗细菌感染相关的不必要的医疗费用并预防 进一步扩大现有的耐药菌株。