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，并且通常具有有限的灵敏度。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著提高了灵敏度并缩短了检测时间，但PCR化学的固有技术限制排除了在单个反应管中进行大规模病原体鉴定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，能够有效的样品制备，随后，细菌病原体的高灵敏度和多重检测。拟议的平台有几个独特的特点。它利用多色光束共聚焦荧光光谱技术，使敏感和多重检测的传染性病原体。此外，该系统还集成了一个微流控芯片，该芯片使用磁致动液滴进行DNA分离，探针-靶杂交和单分子检测。液滴微流体平台允许以无泵和无阀的方式进行流体操纵，而不需要繁琐地集成流体部件和管道，从而大大提高了所提出的病原体检测系统的易用性和通量。 该项目将产生广泛的社会影响。拟议的生物传感系统将使高灵敏度，快速和广泛的检测传染性病原体。由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有关。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挽救生命至关重要。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。高中生将被邀请在PI进行研究？s实验室。此外，实验装置的完成将改善约翰霍普金斯大学微/纳米/生物科学和工程领域的培训基础设施。

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.