Barcoded Nanowires for Multiplexed Clinical Diagnostics

Project: Research project

Project Details

Description

We propose to develop rapid multiplexed assays for the diagnosis of respiratory pathogens in clinical
samples. Many viral and bacterial infections cause respiratory illnesses with very similar symptoms, thus,
timely and accurate pathogen identification is important for patient recovery and public health monitoring.
Current methods include direct immunofluorescence and cell culture, both of which currently require
individual tests for each pathogen. We will use barcoded metallic nanowires as encoded supports for two
types of multiplexed diagnostic assays, which will be tested on clinical samples and ultimately transitioned
into a clinical setting during the course of the proposed work.
Aim 1. Self quenching nanowire beacons. We will take advantage of the electromagnetic effects near metal
surfaces (quenching, enhancement, and alteration of fluorescence lifetimes), to produce nanowire beacon
probes that enable many assays to be conducted simultaneously in the same sample. The resulting
multiplexible, "closed tube" nucleic acid detection assays will require no target labeling or washing, nor any
special instrumentation beyond a conventional fluorescence optical microscope, and are therefore amenable
to point-of-contact assays such as clinical diagnostics. We propose here to develop a 17-plexed assay for
respiratory pathogens.
Aim 2. Simultaneous multiplexed amplification and immobilization for pathogen detection. For viruses shed
at very low levels, amplification (by growing in culture or by PCR) is necessary for detection. We will
overcome current limitations of viral detection by existing methods by introducing multiplexed amplification
and immobilization onto barcoded nanowires (MAIN). To accomplish this we will develop robust surface
attachment chemistry that can withstand thermocycling and will provide for minimal steric hindrance of
hybridization and extension, and will then use these advances to simultaneously direct all PCR products of
interest onto different, visually identifiable nanowire supports for sensitive pathogen detection.
StatusFinished
Effective start/end date7/1/028/31/10

Funding

  • National Institute of Biomedical Imaging and Bioengineering: $295,908.00
  • National Institute of Biomedical Imaging and Bioengineering: $296,486.00
  • National Institute of Biomedical Imaging and Bioengineering: $274,177.00
  • National Institute of Biomedical Imaging and Bioengineering: $185,616.00
  • National Institute of Biomedical Imaging and Bioengineering: $274,455.00
  • National Institute of Biomedical Imaging and Bioengineering: $192,237.00

Fingerprint

Explore the research topics touched on by this project. These labels are generated based on the underlying awards/grants. Together they form a unique fingerprint.