Grantee Research Project Results
Final Report: Handheld FRET-Aptamer Sensor for Chem-Bio Threats
EPA Contract Number: EPD11033Title: Handheld FRET-Aptamer Sensor for Chem-Bio Threats
Investigators: Bruno, John G.
Small Business: Operational Technologies Corporation
EPA Contact: Richards, April
Phase: I
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)
Description:
Operational Technologies Corporation (OpTech) set out in Phase I to expand its patented fluorescence resonance energy transfer (FRET)-DNA aptamer technology (U.S. Patent No. 7,906,280) in the area of chemical and biological threat agent detection. In particular, OpTech used its existing methylphosphonic acid (MPA) aptamer FRET systems to optimize detection of certain organophosphorus (OP) nerve agent simulants using its commercial off-the-shelf (COTS) handheld fluorometer. OpTech also developed new DNA aptamer sequences to bind and detect whole B. anthracis (nonpathogenic Sterne strain) spores and terminal anthrose sugar as well as botulinum neurotoxins (BoNT) A, B and E for rapid FRET detection using the handheld fluorometer.
Summary/Accomplishments (Outputs/Outcomes):
- Developed 280 new patent-pending candidate aptamer DNA sequences against anthrose, whole B. anthracis spores, and the heavy and light chains of botulinum toxins A, B and E.
- Developed an end-labeled aptamer beacon for detection of MPA and some OP agents.
- Developed an aptamer beacon for BoNT A light chain detection to low ng/mL levels in buffer with good specificity.
- Demonstrated drying and rehydration of the BoNT A aptamer beacon without loss of the lower limit of detection.
- Developed two aptamer beacons and intrachain FRET aptamers for low-level detection of Bacillus spores. These FRET aptamers were unable to discriminate B. anthracis from B. cereus or B. thuringiensis, but they could still have value for onsite presumptive testing of unknown powders.
Conclusions:
OpTech has expanded its FRET-aptamer assay technology in the area of chemical and biological defense and homeland security to a point where it soon could be marketable to the first responder community. More work remains, but clearly solid proof-of-principle has been obtained, and OpTech will pursue partnering with established companies in this arena.
Commercialization: OpTech intends to partner with one of the potential sensor companies identified in the Foresight Science and Technology assessment report. OpTech also eventually will demonstrate and market the technology at tradeshows to initiate sales and use sales channels of larger vendor partners.
Journal Articles on this Report : 5 Displayed | Download in RIS Format
| Other project views: | All 5 publications | 5 publications in selected types | All 5 journal articles |
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Bruno JG, Carrillo MP, Phillips T, Vail NK, Hanson D. Competitive FRET-aptamer-based detection of methylphosphonic acid, a common nerve agent metabolite. Journal of Fluorescence 2008;18(5):867-876. |
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Bruno JG, Carrillo MP, Phillips T, Andrews CJ. A novel screening method for competitive FRET-aptamers applied to E. coli assay development. Journal of Fluorescence 2010;20(6):1211-1223. |
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Bruno JG, Zuniga MA, Carrillo MP, Phillips T. Development of naturally selected and molecularly engineered intrachain and competitive FRET-aptamers and aptamer beacons. Combinational Chemistry & High Throughput Screening 2011;14(7):622-630. |
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Bruno JG, Carrillo MP, Phillips T, Hanson D, Bohmann JA. DNA aptamer beacon assay for C-telopeptide and handheld fluorometer to monitor bone resorption. Journal of Fluorescence 2011;21(5):2021-2033. |
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Bruno JG, Carrillo MP, Phillips T, Edge A. Serum inverts and improves the fluorescence response of an aptamer beacon to various vitamin D analytes. Luminescence 2012;27(1):51-58. |
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Supplemental Keywords:
handheld fluorometer, anthracis, DNA aptamer, fluorescence resonance energy transfer (FRET)-DNA aptamer, bioterrorism, biological threat, anthrose, B. anthracis spores, botulinum toxin A, botullinum toxin B, botullinum toxin E.The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.