Handheld FRET-Aptamer Sensor for Chem-Bio ThreatsEPA Contract Number: EPD11033
Title: Handheld FRET-Aptamer Sensor for Chem-Bio Threats
Investigators: Bruno, John G.
Small Business: Operational Technologies Corporation
EPA Contact: Manager, SBIR Program
Project Period: March 1, 2011 through August 31, 2011
Project Amount: $79,984
RFA: Small Business Innovation Research (SBIR) - Phase I (2011) RFA Text | Recipients Lists
Research Category: SBIR - Homeland Security , Small Business Innovation Research (SBIR)
Operational Technologies Corporation (OpTech) proposes to couple the ultrasensitivity of fluorescence-based detection with the high selectivity of synthetic DNA aptamers as replacements for antibodies to detect chemical and biological (CB) terrorism threats in 10 minutes or less from a variety of swabbed surfaces and liquid matrices. OpTech has previously provided strong proof-of-concept for a rapid one-step (“bind and detect”) Fluorescence Resonance Energy Transfer (FRET)-aptamer assay for foot-and-mouth disease virus (J. Biomolec. Techniques, 2008), a nerve gas core molecule (J. Fluorescence, 2008) as well as collagen breakdown peptides and calcidiol to monitor bone metabolism in astronauts (J. Clin. Ligand Assay, 2007). This patent-pending FRET-aptamer assay technology can now be applied in lyophilized form in plastic cuvettes to detect a variety of CB analytes onsite with low ng/ml sensitivity using a commercial off-the-shelf (COTS) handheld and battery-operated fluorometer. In Phase 1, OpTech will screen its existing methylphosphonic acid (MPA; nerve agent core), soman derivative, and Botulinum toxin A (BoNT A) candidate DNA aptamer sequences for FRET assay potential. OpTech also will develop aptamers to “anthrose” surface glycoprotein terminal sugar for detection of Bacillus anthracis spores. In Phase 2, OpTech will fully develop and lyophilize the selected FRET-aptamer assays in plastic cuvettes for use with the sensitive COTS handheld fluorometer for facile, rapid, and cost-effective detection of MPA, sarin or soman, BoNTs, and anthrax spores. OpTech also will expand its repertoire of assays in Phase 2 to include more CB threat agents (viral and bacterial agents of interest).
The primary market for OpTech’s initial CB FRET-aptamer assays will be federal and state first responders as well as military personnel. Clearly, however, the one-step FRET-aptamer assay approach coupled to a handheld COTS fluorometer constitutes a platform technology with broad applications in clinical diagnostics, veterinary diagnostics, food safety testing, and environmental and agricultural monitoring.