Low-cost portable system for the rapid detection and drug resistance profiling of Tuberculosis in low resource environments.

低成本便携式系统，用于在资源匮乏的环境中快速检测结核病并进行耐药性分析。

• 批准号: 9336096
• 负责人: Stephen O'Connor
• 金额: $ 29.01万
• 依托单位: IMPEDX DIAGNOSTICS, INC.
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9336096
• 关键词: Antibiotics; Bacteria; Binding; Buffers; Cell membrane; Cells; Cessation of life; Characteristics; Culture Media; Decontamination; Drug Formulations; Drug resistance; Drug resistance in tuberculosis; Electrodes; Environment; Ethambutol; Formulation; Fostering; Freeze Drying; Genus Mycobacterium; Goals; Growth; Hour; Incidence; Knowledge; Liquid substance; Magnetic nanoparticles; Magnetism; Mechanics; Methods; Michigan; Microfluidics; Missouri; Molecular; Monitor; Multidrug-Resistant Tuberculosis; Mycobacterium smegmatis; Mycobacterium tuberculosis; Organism; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Phase; Polysaccharides; Population; Predisposition; Preparation; Printing; Process; Proliferating; Protocols documentation; Pseudomonas; Public Health; Pyrazinamide; Recovery; Resistance; Resistance profile; Resources; Rifampin; Sampling; Spectrum Analysis; Sputum; Streptococcus; Stress; Surface; Suspensions; System; Techniques; Technology; Testing; Time; Tuberculosis; Universities; Work; World Health Organization; aptamer; bactericide; combat; commensal microbes; cost; design; drug candidate; drug testing; electric impedance; global health; instrument; interest; isoniazid; killings; lithography; mortality; nanoparticle; particle; pathogen; portability; prototype; public health relevance; rapid detection; receptor; sample collection; transmission process; tuberculosis drugs

Tuberculosis (TB) is one of the major public-health challenges in the world today, with an estimated 1.5 million deaths in 2013. More alarmingly, the incidence of multi-drug resistant (MDR) TB is rapidly increasing, especially in low resource environments. The goal of this project is to develop an inexpensive portable system and method using which it will be possible to both detect the presence of low loads of Mycobacterium tuberculosis (Mtb) in sputum samples AND perform Drug Susceptibility Testing (DST) for 12 drug formulations: all within 3 days of sample collection. The proposed approach involves (a) collecting all the ~1000 Mtb cells present in 2-10 ml of sputum sample into a small (200 ul) of buffer (growth medium) during the standard decontamination process using a set of super-paramagnetic nanoparticles (MNPs) functionalized on the surface with amino-polysaccharide moieties that bind to bacterial cells (b) dispensing the 200ul suspension into 16 micro-wells, each containing different formulations of freeze-dried growth media and/or drugs of interest (c)monitor cells in each well using micro-channel Electrical Impedance Spectroscopy (m-EIS). m-EIS reveals in real time if cells are growing, dying, or rendered static in the presence of antibiotic. The first 2 steps (pre-concentration using MNPs and loading into microfluidic channel) will take less than 1 hour, and the monitoring of death/growth/stasis will take 3 days. In Phase I, we will demonstrate proof-of-concept for the above approach by (1) verifying the efficacy of our sample preparation / pre-concentration technique involving MNPs, (2) establishing the ability of the m-EIS method to record in real time, the death, proliferation, or statis of mycobacteria in the presence of candidate drug formulations, and (3) designing a prototype cassette containing 16 wells, and fabricating the same using 3D printing (stereolithography). In addition, we will (4) propose a detailed design (electrical, mechanical and fluidic) of the automated system (disposables and non- disposable hardware) that we will build in Phase II. Being able to detect the presence of low loads of mycobacteria and gaining knowledge about the drug resistance profile of a particular isolate in 3 days will be a “game changer” in our efforts to combat the spread of TB (esp. MDR TB). It will lead to better outcomes for patients (less mortality, quicker recovery), and also better public health (less transmission to others and less fostering of drug resistance in populations).

结核病（TB）是当今世界面临的主要公共卫生挑战之一，据估计， 1.5 2013年死亡人数。更令人担忧的是，耐多药结核病的发病率正在迅速增加，特别是在资源匮乏的环境中。 本项目的目标是开发一种廉价的便携式系统和方法，使用该系统和方法可以检测痰液中低负荷结核分枝杆菌（Mtb）的存在 样品，并对12种药物制剂进行药物敏感性试验（DST）：均在样品采集后3天内进行。 所提出的方法包括（a）将存在于2-10 ml痰样品中的所有~1000个Mtb细胞收集到少量（200 μ l）缓冲液中在标准去污过程期间，使用一组在表面上用结合细菌细胞的氨基-多糖部分官能化的超顺磁性纳米颗粒（MNP）（B）将200 μ l悬浮液分配到16 μ l微-纳米颗粒（生长培养基）中。威尔斯孔，每个孔含有冷冻干燥的生长培养基和/或感兴趣的药物的不同制剂（c）使用微通道电阻抗谱（m-EIS）监测每个孔中的细胞。m-EIS能真实的显示细胞在抗生素存在下是生长、死亡还是静止。前两个步骤（使用MNP预浓缩和加载到微流体通道中）将花费不到1小时，并且死亡/生长/停滞的监测将花费3天。 在I期，我们将通过以下方式证明上述方法的概念验证：（1）验证我们涉及MNP的样品制备/预浓缩技术的功效，（2）建立m-EIS方法在候选药物制剂存在下真实的时间内记录分枝杆菌死亡、增殖或静止的能力，以及（3）设计包含16个威尔斯孔的原型盒，并使用3D打印（立体光刻）制造该材料。此外，我们将（4）提出我们将在第二阶段建立的自动化系统（一次性和非一次性硬件）的详细设计（电气、机械和流体）。 能够在3天内检测到低负荷分枝杆菌的存在并获得有关特定分离株的耐药性特征的知识，将是我们抗击结核病（特别是耐多药结核病）传播的努力中的“游戏规则改变者”。它将为患者带来更好的结果（死亡率更低，康复更快），也将改善公共卫生（减少向他人传播，减少在人群中培养耐药性）。