Integrated Single Molecule Color Coding System for Multiplexed Detection of Pathogens

Johns Hopkins University, Baltimore MD

Investigators

Abstract

Integrated Single Molecule Color Coding System for Multiplexed Detection of Pathogens Among 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.