SBIR-STTR Award

Recent developments in genetically engineered bacteria have opened tremendous opportunities for the Department of Energy and industry in the detoxification of hazardous waste sites. While the benefits are enormous, the resulting bacterial species are not natural to the ecosystem and specific subspecies must be continuously monitored in the field. Monitoring instruments should have detection limits as low as tens of organisms per liter and should have detection times of several minutes. This project will evaluate the ability of piezoelectric detectors to provide the requisite stability, sensitivity and ruggedness for field-based bacterial detection and speciation. Three essential steps will be addressed in Phase I: the selection of the piezoelectric sensor geometry, the selection of an attachment chemistry and the demonstration of sensor performance. The Phase I effort will employ sample injection of crude polymerase chain reaction (PCR) product to demonstrate detection. Phase II will focus on engineering refinement of the system components, integration of sample collection, preconcentration and amplification reactor subsystems with the detector and the evaluation of sensor arrays. Phase II will result in a prototype of a complete system with detect limits of tens of organisms per liter.

Commercial Applications and Other Benefits as described by the awardee:
This project is expected to result in a low cost, simple, sensitive and selective field portable sensor with applications in environmental monitoring, food processing, clinical diagnostics and medical research. The instrument will address Federal, state and municipal requirements, as well as industrial (e.g. agriculture, food processing and fisheries) requirements for bacterial detection and speciation.

The detoxification of hazardous waste sites is a serious, widespread, and ongoing problem. Recent developments in genetically engineered bacteria have allowed new approaches to detecting, monitoring, and eliminating dangerous materials from our air and aquifers. Remediation efforts require the use of an instrument that can autonomously identify, observe, and analyze specific subspecies of bacteria. The device should have detection limits as low as tens of organisms per liter and should have detection times as short as several minutes. This project will develop instrumentation based on piezoelectric detectors that provide the requisite stability, sensitivity, and ruggedness for field-based bacterial detection and speciation. Phase I demonstrated suitable surface transverse wave sensor geometries, verified attachment chemistries for DNA probes, and demonstrated of a viable piezoelectric sensor for E. coli DNA. Phase II will address three essential steps: development of the piezoelectric sensing structure, development of the optimum attachment chemistry, and demonstration of sensor performance in real situations. The Phase II effort will ultimately integrate polymerase chain reaction amplification of deoxyribonucleic acid with the piezoelectric sensor. Temperature drift, aging, and other difficulties will be overcome by intelligent instrumentation. This approach will also minimize the time needed to obtain a measurement and will provide semiquantitative analysis. Phase II will result in a prototype of a complete system with detection limits of tens of organisms per liter. Commercial Applications and other Benefits as described by the awardee. A low cost, simple, sensitive, and selective field-portable sensor should find application in environmental monitoring, food processing, clinical diagnostics, and medical research. The resultant device will be able to verify compliance with federal, state, and municipal mandates, as well as industrial (e.g., agriculture, food processing, and fisheries) requirements for bacterial detection and speciation.