SBIR-STTR Award

Current geopolitical strategies require effective warfare countermeasures to protect U.S. forces against biological and chemical threats. Today's molecular biology and immunological detection technologies cannot be used to produce automated biodetectors useable in the battlefield. New, improved detection technologies are needed to minimize the impact of chemical and biological weapons on Army personnel and provide a means to construct automated biodetectors that can identify a very few particles of any hazardous agent, regardless of interferant background. The final objective of the work proposed is to develop a highly sensitive RNA probe methodology to identify pathogenic bacteria and viruses. The methodology will underlay the foundation of RNA based tests that will be easy to perform as a simple enzymatic reaction at 37 degrees C under isothermal conditions in a single test tube format and can identify less than 100 target molecules in a specimen. Because of its extreme simplicity, the test could ultimately be incorporated into a small, portable, personal devicuseable in battlefield environments without special training. This technology goes beyond normal evolutionary development approaches because it breaks the existing paradigms of the diagnostic industry by demonstrating the technical feasibility of detecting a non-nucleic acid target using a nucleic acid detector system.

Keywords:
Recombinant Rna Detectors, 0-Beta Replicase, Non-Nucleic Acid Targets, Probe Amplification System, F

We propose to design and construct a prototype device for rapid microorganiam identification using RNA probe methodology. The device will contain a biochip cartridge with molecular detection systems for selected biomarkers, and will employ amplifiable RNA probes to identify simultaneously a model group of RNA and protein biomarkers representing bacteria (Bacillus species) and RNA viral agents (HIV and HCV). The biochips will register the presence of biomarkers specific for individual bacterial species, bacterial spores or viral agents in a specimen as an amplified RNA product that will be generated upon binding to the targeted biomarkers. The detector device prototype will work in connection with a specimen preparation cartridge of our design that processes environmentally-obtained dirty samples. The prototype and sample preparation devices could be miniaturized and will operate efficiently under extreme environmental conditions in any locale. The proposed device will feature proprietary detection technology tested successfully in Phase I and will be used to further validate the technology's ability to identify targets of any nature, including nucleic acid, proteins, toxins, small molecules and bioregulators. Prior research indicates that our new system will overcome the limitations inherent in other available detection systems, such as immunodiagnostics and PCR. Exploiting this methodology will have a significant impact on areas of military and civilian importance. The proposed technology will serve as a common platform to detect biological warfare agents and medicinally important compounds that so far have been our of reach for routinely used detection technologies due to their inherent common drawbacks, such as high cost, complexity and time required to perform, and more specific limitations, such as restructions in application (nucleic acid technology) or inadequate sensitivity (immunodiagnostics).

Keywords:
Recombinant Rna Detectors, 0-Beta Replicase, Non-Nucleic Acid Targets, Probe Amplification System, F