Understand and Detect Sepsis: Pathogen Isolation, Biochemistry Assay, and Optofluidic Sensing

了解和检测脓毒症：病原体分离、生物化学检测和光流控传感

• 批准号: 10671479
• 负责人: Ke Du
• 金额: $ 36.67万
• 依托单位: UNIVERSITY OF CALIFORNIA RIVERSIDE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10671479
• 关键词: Address; Antimicrobial Resistance; Bacteria; Bacterial Infections; Biochemistry; Biological Assay; Biological Markers; Biomedical Research; Blood specimen; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Communities; Consumption; Detection; Diagnosis; Disease; Early treatment; Emergency Situation; Engineering; Fluorescence; Foundations; Goals; Home; Interdisciplinary Study; Label; Laboratories; Life; Liquid substance; Malignant Neoplasms; Methods; Microfluidic Microchips; Molecular Diagnosis; Nanostructures; Patients; Persons; Postdoctoral Fellow; Preparation; Prevention; Property; Public Health; Publishing; Research; Research Personnel; Resistance; Retrieval; Risk; Sampling; Sensitivity and Specificity; Sepsis; Septic Shock; Students; Technology; Testing; Tissues; Virus; Virus Diseases; Vision; Whole Blood; Work; accurate diagnosis; advanced disease; antimicrobial; bacterial resistance; disease diagnosis; doctoral student; interdisciplinary approach; journal article; miniaturize; mortality; nanodevice; nanomaterials; nervous system disorder; nucleic acid detection; pathogen; programs; rapid diagnosis; response; small molecule; undergraduate student; waveguide; whole blood bacteria

Project Summary Background: Sepsis is a life-threatening emergency, normally caused by the body’s response to a bacterial infection. Without early treatment, sepsis can lead to septic shock with approximately 50% mortality rate. Rapid and accurate diagnosis of sepsis is the key to decrease the mortality rate. However, there is no global standard for sepsis testing due to the inadequate sensitivity and specificity of the current technologies. The PI has the ambition to address the critical and far-reaching bottlenecks specifically of concern in sepsis testing: 1) a rapid and simple method to isolate and concentrate bacteria from whole blood sample, 2) a sensitive and one-step CRISPR microfluidic chip to detect the nucleic acid biomarkers of the pathogens, and 3) a multiplexing and miniaturized optofluidic waveguide platform to enhance the fluorescence based detection. Overview of the laboratory: The PI established his own lab at RIT in 2018. With the overwhelm start-up support by the home department, the PI is leading an active and interdisciplinary research group with 1 postdoc researcher, 4 Ph.D. students, 2 Master students, and a couple of undergraduate students. Within 2 years, the lab has published ~10 journal articles in the fields of bacteria/virus isolation, CRISPR biochemistry assay, and optofluidic sensing. Exploiting interdisciplinary approach, the lab is working on technologies to quickly identify antimicrobial resistant bacteria in whole blood sample. As more strains become resistant to available therapies, the risk for people developing life-threatening sepsis is increasing. The research topic we are working on will be the key for clinicians to provide quick clinical decisions and increase the chances of survivals. In addition, the technologies developed in our lab will also lay the foundation for the diagnosis and treatment of many different kinds of diseases such as cancer, viral infection, and neurological diseases. Goals for the next 5 years: Our first goal is to develop a fully automated nanodevice that can collect and concentrate bacteria from whole blood with a retrieval efficiency of 99% and a concentration factor of 10,000. We will begin with spiked sample and then proceed to clinical sample. Leveraging the unique properties of nanomaterials and nanostructures, the second goal is to develop a one-step and isothermal CRISPR chip for low concentration (1 CFU/mL) bacteria detection without front end target amplification. Towards the end of the fifth year, we will integrate the sample preparation chip and the CRISPR detection chip as a single and compact unit for the testing of clinical samples. The third goal is to develop a liquid-core, superhydrophobic nanostructure cladding waveguide platform for multiplexing bacteria detection. The high fluorescence collection efficiency will enable sensitive sepsis detection with microliter level sample consumption. Overall vision of the research program: The technologies we are developing will have a broad impact to the biomedical research communities to understand and engineering small molecules, cells, and tissues. The proposed work will also advance disease diagnosis, prevention, and treatment.

项目摘要 背景：脓毒症是一种危及生命的紧急情况，通常由身体对细菌的反应引起 感染。如果不及早治疗，脓毒症可导致感染性休克，死亡率约为50%。快速 而准确诊断脓毒症是降低病死率的关键。然而，没有一个全球标准。 由于目前技术的敏感度和特异度不够高，导致败血症检测。少年派有 雄心勃勃地解决败血症检测中特别令人担忧的关键和深远的瓶颈：1)快速 以及从全血样本中分离和浓缩细菌的简单方法，2)灵敏的一步法 CRISPR微流控芯片，用于检测病原体的核酸生物标志物；以及3)多路复用器和 小型化光流控波导平台，增强基于荧光的检测。 实验室概述：2018年，PI在RIT建立了自己的实验室。有了压倒性的创业支持 在内政部，PI正在领导一个积极的跨学科研究小组，拥有1名博士后 研究员，博士生4人，硕士生2人，本科生2人。在两年内， Lab在细菌/病毒分离、CRISPR生化分析和 光流感测。利用跨学科的方法，该实验室正在研究技术，以快速识别 全血标本中存在耐药细菌。随着越来越多的菌株对可用的治疗方法产生抗药性， 人们患上危及生命的败血症的风险正在增加。我们正在进行的研究课题将是 对于临床医生来说，提供快速的临床决策和增加生存机会的关键。此外， 我们实验室开发的技术也将为许多不同疾病的诊断和治疗奠定基础 各种疾病，如癌症、病毒感染和神经系统疾病。 未来5年的目标：我们的第一个目标是开发一种完全自动化的纳米设备，可以收集和 从全血中浓缩细菌，回收率为99%，浓缩系数为10,000。 我们将从添加样本开始，然后进行临床样本。利用的独特属性 纳米材料和纳米结构，第二个目标是开发一种一步等温CRISPR芯片，用于 低浓度(1 cfu/m L)细菌检测，无需前端靶标扩增。在接近尾声的时候 第五年，我们将把样品制备芯片和CRISPR检测芯片整合为单一的紧凑型芯片 临床样本检测单位。第三个目标是开发一种液核超疏水纳米结构 用于多路细菌检测的包层波导台。高的荧光收集效率将 能够以微升水平的样本消耗量实现敏感的败血症检测。 研究计划的总体愿景：我们正在开发的技术将对 生物医学研究社区，以了解和设计小分子、细胞和组织。这个 拟议的工作还将促进疾病的诊断、预防和治疗。