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: 10028772
• 负责人: Peter Yunker
• 金额: $ 30.24万
• 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/10028772
• 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; Diagnostic; Dyes; Infection; Interferometry; Intermediate resistance; Knowledge; Laboratories; Laboratory Research; Malignant Neoplasms; Manuals; Measurement; Measures; Medical; Minor; Modern Medicine; Phenotype; Population; Predisposition; Property; 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）强调了抗生素耐药性是最伟大的医学之一 21世纪的挑战。没有新的抗生素，可以预测 到2050年，全球每年1000万例死亡（超过癌症）。这些感染被毒成否定 依赖抗生素的现代医学的许多进步。那，根据 细菌敏感性将带来最大的治疗益处。 绝大多数未被发现的一种阻力是异性抗性，其中较小的亚群 细菌细胞在表型上具有抗性。这些抗性细胞在存在抗生素的情况下迅速扩展 因此会导致治疗失败。由于异质抗性很常见（> 25％的抗生素 - 细菌组合），所以 检测异性抗性对于解决抗生素耐药性日益严重的危机至关重要。确实，最近是 证明了杂质的知识可用于指导成功的联合疗法，并且 因此，甚至可以治疗抗泛抗菌细菌。 不幸的是，临床敏感性测试缺乏鉴定异质抗衡的解决方案，实验室测试 因为异质剂是耗时且缓慢的。此外，目前下的下一代敏感性测试 开发主要集中在对少数细胞上进行的快速诊断，进行检测 不可能的稀有细胞。开发新的敏感性测试使人的要求更加复杂 临床微生物学实验室；要采用，易感性测试必须需要最少的体力劳动，工作 坚固且价格便宜的 - 通常与高灵敏度不一致的特性。基于这些问题， 广泛同意需要一种新的技术方法。 在这里开发了使用干扰来测量细菌的新的，更敏感的敏感性测试 提出了人口地形。使用干涉测量地形没有优先 抗生素敏感性测试，生物医学研究中的优先次数很少。但是，它通常用于 物理科学既快速，便宜，健壮，不需要染料或污渍，并且提供超分辨率 地形的测量。初步结果表明，干扰具有解决能力 迅速检测出异质剂并将其与易感性区分开。我们基于干涉法的方法是 研究实验室无法检测到异质抗性的临床敏感性测试的实质性改善 可以检测出异质剂但临床应用太慢的测试和传统的地形成像 方法，例如共聚焦显微镜，昂贵，缺乏区分的分辨能力 来自敏感性的异性含量。建立我们的诺言初步结果，该提案将决定 将细菌形态与死亡和繁殖有关的基本生物物理学。这项工作至关重要 在广泛的临床相关细菌和抗生素中，将地形转化为敏感性谱。