High-Throughput, Massively Parallel Antimicrobial Resistance Surveillance Using Drop-Based Microfluidics

使用基于液滴的微流体进行高通量、大规模并行抗菌药物耐药性监测

• 批准号: 10218860
• 负责人: Connie B Chang
• 金额: $ 21.6万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-22 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10218860
• 关键词: Agar; Antibiotic Resistance; Antibiotic Therapy; Antibiotic susceptibility; Antibiotics; Antimicrobial Resistance; Antimicrobial susceptibility; Automation; Bacteria; Bacterial Infections; Bar Codes; Biological Assay; Caliber; Cell Proliferation; Cells; Cessation of life; Clinical; Combined Antibiotics; Communicable Diseases; Detection; Devices; Diagnosis; Diagnostic; Drops; Early Diagnosis; Encapsulated; Engineering; Equipment; Fluorescence; Goals; Growth; Hour; Individual; Infection; Laboratories; Libraries; Life; Liquid substance; Methods; Microfluidic Microchips; Microfluidics; Miniaturization; Minimum Inhibitory Concentration measurement; Monitor; Optics; Pharmacotherapy; Phenotype; Predisposition; Pseudomonas aeruginosa; Research; Resistance; Resolution; Sampling; Sepsis; Signal Transduction; Speed; Stains; Technology; Test Result; Testing; Time; Urinary tract infection; Work; bacterial resistance; base; cell growth; combinatorial; cost; design; detection limit; drug standard; effective therapy; high throughput technology; infectious disease treatment; light scattering; microbial; multiplex assay; new technology; novel; operation; optical fiber; pathogen; population based; portability; prevent; rapid detection; rapid test; resistance mechanism; resistant strain; response; screening; single cell analysis

A fundamental challenge in administering effective treatments for infectious diseases caused by bacteria is the rapid identification of antimicrobial resistance and the efficacy of single or combinatorial drug treatments, including the lowest concentrations or combinations of antibiotics required to prevent bacterial growth. We propose to overcome this by engineering an easy-to-use platform based on miniaturization of microbial cultivation using microfluidic drops that can be tagged, tracked, and evaluated in scalable and massively parallel designs. Standard antimicrobial susceptibility testing (AST) platforms typically require 18-72 hours to generate susceptibility results. The primary research objective of this proposal is to apply droplet-based microfluidics to perform rapid AST screening of P. aeruginosa clinical isolates in under four hours. Droplet-based microfluidics is a technology in which picoliter-sized volumes are created and assayed at rates of up to thousands per second. These drops serve as individual microreactors that can contain single cells. The ability to isolate single cells and discrete combinations of antibiotics within picoliter-sized microreactor volumes will allow for rapid and early detection of antibiotic susceptibility with high resolution and fidelity. This method will enable the ability to detect and quantify subpopulations of single cells that are normally below the limit of detection of standard drug assays. To achieve this goal, we have two specific aims: (1) High-throughput minimum inhibitory concentration screening will be performed by multiplexed assaying with a barcoded droplet library, and (2) A microfluidic device with integrated optical fibers will be engineered for sensitive detection of barcode and cell signals, which will enable the platform to be portable and readily adaptable to clinical settings. This novel, scalable technology for high-throughput antimicrobial susceptibility testing in bacteria can greatly decrease the diagnosis time of bacterial infections such as sepsis or urinary-tract infections.

通过细菌引起的有效治疗的有效治疗方法是快速鉴定抗菌耐药性和单一或联合药物治疗的功效，包括预防细菌生长所需的抗生素所需的浓度最低或组合。我们建议通过使用微流体液滴进行微生物培养的微型化，以在可扩展且大量的并行设计中进行标记，跟踪和评估的微流体滴剂来克服这一点。标准抗菌敏感性测试（AST）平台通常需要18-72小时才能产生敏感性结果。该提案的主要研究目标是应用基于液滴的微流体，以在四个小时内对铜绿假单胞菌临床分离株进行快速筛查。基于液滴的微流体学是一项技术，在该技术中，以每秒多达数千次的速度创建和测定了Picoliter尺寸的体积。这些滴剂是可以包含单个细胞的单个微反应器。在Picoliter大小的微反应器体积中分离单个细胞和抗生素的离散组合的能力将允许快速和早期检测抗生素易感性，并具有高分辨率和忠诚度。该方法将使通常低于标准药物测定的限制的单个细胞的亚细胞进行检测和量化亚群。为了实现这一目标，我们有两个具体的目的：（1）将通过与条形码液滴库的多路复用测定进行高通量抑制浓度筛选，并且（2）具有集成光纤的微流体设备将经过精心设计，以进行敏感的测试和电池信号，以启用该平台，以供台式设置，以供台式设置。这种新型的可扩展技术用于细菌中的高通量抗菌敏感性测试可以大大减少细菌感染的诊断时间，例如脓毒症或尿液感染。