Development of a Rapid Antibiotic Susceptibility Test Capable of Detecting Heteroresistance

开发能够检测异抗性的快速抗生素敏感性测试

• 批准号: 10439871
• 负责人: Peter Yunker
• 金额: $ 30.25万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10439871
• 关键词: Address; Adopted; Agreement; Antibiotic Resistance; Antibiotic susceptibility; Antibiotics; Bacteria; Biomedical Research; Biophysics; Cells; Cessation of life; Clinical; Clinical Microbiology; Combined Modality Therapy; Confocal Microscopy; Consumption; Detection; Development; Dyes; Infection; Interferometry; Intermediate resistance; Knowledge; Laboratories; Laboratory Research; Malignant Neoplasms; Manuals; Measurement; Measures; Medical; Minor; Modern Medicine; Phenotype; Population; Predisposition; Property; Rapid diagnostics; Reproduction; Resistance; Resolution; Stains; Testing; Therapeutic; Time; Treatment Failure; Work; World Health Organization; antibiotic resistant infections; bacterial resistance; base; clinical application; clinically relevant; imaging approach; new combination therapies; new technology; next generation; novel strategies; physical science

The World Health Organization (WHO) has highlighted antibiotic resistance as one of the greatest medical challenges of the 21st century. Without new antibiotics, it is predicted that antibiotic resistant infections will cause 10 million annual deaths worldwide by the year 2050 (surpassing cancer). These infections are poised to negate many advances of modern medicine that rely on antibiotics. Thus, choosing effective antibiotics based on bacterial susceptibilities would provide the greatest therapeutic benefit. One form of resistance that is overwhelmingly undetected is heteroresistance, in which a minor subpopulation of bacterial cells is phenotypically resistant. These resistant cells rapidly expand in the presence of an antibiotic and thus can cause treatment failure. As heteroresistance is common (>25% of antibiotic-bacteria combinations), detecting heteroresistance is critical to address the growing crisis of antibiotic resistance. Indeed, it was recently demonstrated that knowledge of heteroresistance can be used to guide successful combination therapy, and can thus even treat pan-resistant bacteria. Unfortunately, clinical susceptibility tests lack the resolution to identify heteroresistance, and the laboratory test for heteroresistance is time consuming and slow. Further, the next-generation susceptibility tests currently under development are largely focused on rapid diagnostics performed on a small number of cells, making detection of rare cells impossible. Developing a new susceptibility test is further complicated by the requirements of the clinical microbiology laboratory; to be adopted, a susceptibility test must require minimal manual labor, work robustly, and be inexpensive – properties that are often at odds with high sensitivity. Based on these issues, there is broad agreement that a new technological approach is required. Here the development of a new, more sensitive susceptibility test using interferometry to measure bacterial population topography is proposed. The use of interferometry to measure topography has no precedence in antibiotic susceptibility testing, and little precedence in biomedical research. However, its commonly used in physical sciences as it is rapid, inexpensive, robust, doesn’t require dyes or stains, and provides super-resolution measurements of topography. Preliminary results demonstrate that interferometry has the resolving power to rapidly detect heteroresistance and distinguish it from susceptibility. Our interferometry-based approach is a substantial improvement over clinical susceptibility tests that cannot detect heteroresistance, research laboratory tests that can detect heteroresistance but are too slow for clinical application, and traditional topographic imaging approaches, e.g. confocal microscopy, that are expensive and lack the resolving power to distinguish heteroresistance from susceptibility. Building off our promising preliminary results, this proposal will determine the underlying biophysics relating bacterial topography to death and reproduction. This work is pivotal to reliably convert topography into susceptibility profiles across a broad range of clinically relevant bacteria and antibiotics.

世界卫生组织(WHO)强调，抗生素耐药性是最大的医疗疾病之一 21世纪的挑战。如果没有新的抗生素，预计抗药性感染将导致 到2050年，全球每年有1000万人死亡(超过癌症)。这些感染可能会抵消 现代医学的许多进步都依赖于抗生素。因此，根据以下因素选择有效的抗生素 细菌敏感性将提供最大的治疗益处。 一种压倒性未被检测到的抗性形式是异源抗性，在这种情况下，一个较小的亚群 细菌细胞是表型耐药的。这些耐药细胞在抗生素的存在下迅速扩张。 从而可能导致治疗失败。由于异源耐药是常见的(25%的抗生素-细菌组合)， 检测异源耐药性对于解决日益严重的抗生素耐药性危机至关重要。事实上，这是最近的事 证明异耐药的知识可以用来指导成功的联合治疗，以及 因此，甚至可以治疗泛耐药细菌。 不幸的是，临床药敏试验缺乏确定异质性耐药的分辨率，而实验室试验 因为异质电阻既耗时又缓慢。此外，目前正在进行的下一代敏感性测试 开发主要集中在对少量细胞执行快速诊断，使检测 稀有细胞是不可能的。开发一种新的药敏试验因 临床微生物学实验室；要被采用，药敏试验必须需要最少的体力劳动 坚固、廉价--这些房产往往与高度敏感的房产格格不入。基于这些问题， 人们普遍认为，需要一种新的技术方法。 这里开发了一种新的、更灵敏的药敏试验，使用干涉法来测量细菌 提出了人口拓扑学的概念。使用干涉测量来测量地形在以下方面是没有先例的 抗生素敏感性测试，在生物医学研究中几乎没有优先地位。然而，它通常用于 物理科学，因为它是快速的，廉价的，坚固的，不需要染料或染色，并提供超分辨率 地形测量。初步结果表明，干涉测量具有分辨能力 快速检测异质电阻，并将其与敏感性区分开来。我们基于干涉测量的方法是一种 研究实验室：对无法检测出异源耐药的临床药敏试验有实质性改进 可以检测到异质电阻但对于临床应用来说太慢的测试，以及传统的地形图成像 昂贵且缺乏分辨能力的方法，如共聚焦显微镜 由敏感性产生的异质电阻。在我们有希望的初步结果的基础上，这项提案将确定 细菌地形与死亡和繁殖有关的基本生物物理学。这项工作对于可靠地 将地形转换为广泛的临床相关细菌和抗生素的药敏图谱。