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: 10669000
• 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/10669000
• 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; clinical application; clinically relevant; imaging approach; improved; new combination therapies; new technology; next generation; novel strategies; physical science; rapid detection; ultra high resolution

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严重的医学问题之一 21世纪的挑战。如果没有新的抗生素，预计抗生素耐药性感染将导致 到 2050 年，全球每年将有 1000 万人死亡（超过癌症）。这些感染有望消除 现代医学的许多进步都依赖于抗生素。因此，根据情况选择有效的抗生素 细菌敏感性将提供最大的治疗益处。 一种绝大多数未被检测到的耐药形式是异质耐药，其中一小部分亚群 的细菌细胞具有表型耐药性。这些耐药细胞在抗生素存在下迅速扩增 从而导致治疗失败。由于异质耐药性很常见（>25% 的抗生素-细菌组合）， 检测异质耐药性对于解决日益严重的抗生素耐药性危机至关重要。确实，最近 证明异质耐药性的知识可用于指导成功的联合治疗，并且 因此甚至可以治疗泛耐药细菌。 不幸的是，临床药敏试验缺乏识别异质抗性的分辨率，而实验室试验 因为异质抗性是耗时且缓慢的。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进行快速诊断，使检测 稀有细胞是不可能的。开发新的敏感性测试因要求而变得更加复杂 临床微生物学实验室；要采用，敏感性测试必须需要最少的体力劳动、工作 坚固耐用且价格低廉——这些特性通常与高灵敏度相矛盾。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、更灵敏的药敏测试，使用干涉测量法来测量细菌 提出了人口地形。使用干涉测量法测量形貌在以下领域中没有先例： 抗生素敏感性测试，在生物医学研究中几乎没有优先权。然而，它通常用于 物理科学，因为它快速、廉价、稳健，不需要染料或染色剂，并且提供超分辨率 地形测量。初步结果表明干涉测量法具有分辨能力 快速检测异质抗性并将其与易感性区分开来。我们基于干涉测量的方法是 与无法检测异质抗性的临床药敏试验相比，研究实验室有显着改善 可以检测异质耐药性但对于临床应用而言速度太慢的测试以及传统的地形成像 方法，例如共焦显微镜，价格昂贵且缺乏分辨能力 来自易感性的异质抗性。基于我们有希望的初步结果，该提案将确定 将细菌形态与死亡和繁殖相关的基础生物物理学。这项工作对于可靠地 将地形图转换为各种临床相关细菌和抗生素的敏感性概况。