Interferometry to understand beta-lactam heteroresistance

干涉测量法了解 β-内酰胺异质抗性

• 批准号: 10798548
• 负责人: Peter Yunker
• 金额: $ 12.65万
• 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/10798548
• 关键词: Address; Adopted; Agreement; Antibiotic Resistance; Antibiotic susceptibility; Antibiotics; Award; Bacteria; Biomedical Research; Biophysics; Cells; Clinical Microbiology; Diagnostic; Dyes; Equipment; Funding; Growth; Hour; Instruction; Interferometry; Intermediate resistance; Laboratories; Manuals; Measurement; Measures; Medical; Minor; Modern Medicine; Parents; Phenotype; Physics; Population; Predisposition; Process; Property; Research; Resistance; Stains; Testing; Therapeutic; Time; Treatment Failure; Work; World Health Organization; beta-Lactam Resistance; beta-Lactams; biophysical analysis; clinically relevant; design; experimental study; flexibility; materials science; new technology; parent grant; ultra high resolution

The World Health Organization (WHO) has highlighted antibiotic resistance as one of the greatest medical challenges of the 21st century. Choosing effective antibiotics is therefore critical, and basing antibiotic selection on bacterial susceptibilities potentially offers the greatest therapeutic benefit. Current antibiotic susceptibility tests are unable to detect a common form of resistance (present in more than 25% of antibiotic-bacteria combinations) known as 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. 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. In the parent grant, we proposed to develop a new susceptibility test using interferometry to measure bacterial population topography. The use of interferometry to measure topography has no precedence in antibiotic susceptibility testing, and little precedence in biomedical research in general. However, it is common in physics and materials science as it is rapid, doesn’t require dyes or stains, and provides super-resolution measurements of topography. Interferometers are inexpensive and robust. In exciting preliminary results, we have demonstrated that with interferometry we can detect heteroresistance in 2 hours, and even distinguish low levels of heteroresistance from susceptibility. In this equipment supplement, we are requesting funds to purchase another interferometer. In the process of developing an interferometry-based susceptibility test, we made a discovery about the biophysics of beta-lactam heteroresistance. In line with the parent grant’s proposed work on measuring and understanding bacterial topography, as well as proposed work on interferometry-based diagnostics, here we propose to investigate how the growth of beta-lactam resistant cells depends on the local concentration of resistant cells. These experiments are necessarily long (each takes at least 24 hours) and drawing conclusions will require testing many different strains against different beta-lactams, and thus require additional equipment. This project addresses fundamental questions about the biophysics of rare resistant cells. Further, as beta- lactams are the most used class of antibiotics, the results of the work proposed here will also hold clinical relevance.

世界卫生组织（WHO）强调，抗生素耐药性是最大的医学威胁之一。 世纪的挑战。因此，选择有效的抗生素至关重要， 对细菌亲和性的影响可能提供最大的治疗益处。当前抗生素敏感性 测试无法检测出一种常见的耐药性（存在于超过25%的寄生细菌中 组合）称为异源耐药性，其中细菌细胞的次要亚群在表型上是 抵抗这些耐药细胞在抗生素存在的情况下迅速扩增，因此可以引起治疗 失败临床微生物学的要求使开发新的药敏试验变得更加复杂 实验室;要被采用，敏感性测试必须需要最少的手工劳动，工作稳健， 廉价--通常与高灵敏度不一致的特性。基于这些问题， 他们认为需要一种新的技术方法。 在母基金中，我们建议开发一种新的药敏试验，使用干涉测量法来测量细菌 人口地形学使用干涉测量术测量地形在抗生素中没有先例 敏感性测试，在一般生物医学研究中几乎没有先例。然而，在物理学中， 和材料科学，因为它是快速的，不需要染料或染色，并提供超分辨率测量 地形学。干涉仪价格低廉且坚固耐用。在令人兴奋的初步结果中，我们证明了 通过干涉测量法，我们可以在2小时内检测到异源抗性，甚至可以区分低水平的 从易感性到异源抗性。 在这一设备补充中，我们要求资金购买另一台干涉仪。过程中 通过开发一种基于干涉仪的药敏试验，我们发现了β-内酰胺的生物物理学 异源抗性根据母基金提出的测量和了解细菌的工作， 地形，以及提出的工作干涉为基础的诊断，在这里，我们建议调查如何 β-内酰胺抗性细胞的生长取决于抗性细胞的局部浓度。这些实验 必须很长（每个至少需要24小时），得出结论将需要测试许多不同的 菌株对不同的β-内酰胺，因此需要额外的设备。 该项目解决了有关罕见耐药细胞生物物理学的基本问题。此外，作为beta- 内酰胺类抗生素是最常用的一类抗生素，本文提出的工作结果也将适用于临床 本案无关