Integrated Single Molecule Color Coding System for Multiplexed Detection of Pathogens

用于病原体多重检测的集成单分子颜色编码系统

• 批准号: 0967375
• 负责人: Jeff Wang
• 金额: $ 34万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0967375&HistoricalAwards=false
• 关键词: Integrated; Single; Molecule; Color; Coding

Integrated Single Molecule Color Coding System for Multiplexed Detection of PathogensAmong the various methods currently used in clinical laboratories for the detection of bacterial pathogens, the cell culture-based method remains the gold standard. However, this method may require 24 hours or longer obtaining results and often has limited sensitivity. Although PCR-based approaches to pathogen detection have significantly improved sensitivity and shortened detection times as compared to conventional culture-based systems, the inherent technical limitation of PCR chemistry precludes sufficient multiplexing capacity for large-scale pathogen identification and characterization in a single reaction tube. The project focuses on developing an integrated biosensing system capable of efficient sample preparation and, subsequently, highly sensitive and multiplexed detection of bacterial pathogens. The proposed platform has several unique features. It utilizes the multi-color beam shaped confocal fluorescence spectroscopic technology to enable sensitive and multiplexed detection of infectious agents. In addition, the system incorporates a microfluidic chip that uses magnet-actuated droplets for DNA isolation, probe-target hybridization and single molecule detection via beam-shaped confocal spectroscopy. The droplet microfluidic platform allows fluidic manipulations in a pump-less and valve-less manner without the need for tedious integration of fluidic components and tubing, thereby greatly enhancing the ease-of-use and throughput of the proposed pathogen detection system. The proposed project will have immediate broad and social impact. The proposed biosensing system will enable highly sensitive, rapid, and broad-based detection of infectious pathogens. Diseases caused by infectious pathogens are major causes of death, disability, and social and economic disruption for millions of people globally. Diagnosis forms the basis for accurately treating diseases, and timely therapeutic intervention is associated with improved clinical outcome and prognosis. Thus, the development of a biosensing platform capable of rapid and accurate diagnosis of infectious pathogens is critical to saving lives. In addition, this research will provide excellent opportunities for cross-disciplinary training for K-12, undergraduate and graduate students. High school students will be invited to perform research in the PI?s lab. Furthermore, the completion of the experimental setup will improve the infrastructure for training in the area of micro/nano/bio science and engineering at the Johns Hopkins University.

用于病原体多重检测的集成单分子颜色编码系统在临床实验室目前用于检测细菌病原体的各种方法中，基于细胞培养的方法仍然是金标准。然而，这种方法可能需要 24 小时或更长时间才能获得结果，并且灵敏度通常有限。尽管与传统的基于培养的系统相比，基于 PCR 的病原体检测方法显着提高了灵敏度并缩短了检测时间，但 PCR 化学固有的技术限制阻碍了在单个反应管中进行大规模病原体识别和表征的足够的多重能力。该项目的重点是开发一个集成的生物传感系统，能够有效地制备样品，并随后对细菌病原体进行高灵敏度和多重检测。所提议的平台有几个独特的功能。它利用多色光束形状共焦荧光光谱技术来实现对传染性病原体的灵敏和多重检测。此外，该系统还集成了一个微流控芯片，该芯片使用磁力驱动的液滴进行 DNA 分离、探针-靶标杂交以及通过束形共焦光谱进行单分子检测。液滴微流体平台允许以无泵和无阀的方式进行流体操作，而不需要流体组件和管道的繁琐集成，从而大大提高了所提出的病原体检测系统的易用性和吞吐量。 拟议的项目将立即产生广泛的社会影响。所提出的生物传感系统将能够对传染性病原体进行高度灵敏、快速和广泛的检测。传染性病原体引起的疾病是全球数百万人死亡、残疾以及社会和经济混乱的主要原因。诊断是准确治疗疾病的基础，及时的治疗干预与改善临床结果和预后相关。因此，开发能够快速准确诊断传染性病原体的生物传感平台对于拯救生命至关重要。此外，这项研究将为K-12、本科生和研究生提供跨学科培训的绝佳机会。高中生将被邀请到 PI 的实验室进行研究。此外，实验装置的完成将改善约翰·霍普金斯大学微/纳米/生物科学与工程领域的培训基础设施。