Integrated Single Molecule Color Coding System for Multiplexed Detection of Pathogens

在目前临床实验室用于细菌性病原体检测的各种方法中，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，而且通常灵敏度有限。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著提高了灵敏度，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鉴定和表征的足够多路处理能力。该项目的重点是开发一种综合生物传感系统，能够有效地制备样品，随后对细菌病原体进行高度敏感和多重检测。提出的平台有几个独特的特点。它利用多色光束形共聚焦荧光光谱技术，使敏感和多路检测感染因子。此外，该系统还集成了一个微流控芯片，该芯片使用磁体驱动液滴进行DNA分离、探针-靶标杂交和单分子检测，通过光束形共聚焦光谱。液滴微流控平台允许以无泵和无阀的方式进行流体操作，而无需将流体元件和管道进行繁琐的集成，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和社会影响。提出的生物传感系统将能够高度敏感，快速和广泛的检测感染性病原体。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中断的主要原因。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感染性病原体的生物传感平台对于挽救生命至关重要。此外，本研究将为K-12，本科生和研究生的跨学科培训提供良好的机会。高中学生将被邀请到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/生物科学与工程领域培训的基础设施。

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.