Amplification of Molecular Signals

The goal of the proposed research is to develop a highly sensitive and specific amplification system that can be coupled to any pathogen detection and quantification unit for sensitive real-time detection of biological warfare agents. The system is builtaround a dual-enzyme substrate-recycling electrochemical amplification system coupled to an immunoassay detection technology that has already been proven at a benchtop scale.The specific aims of this effort are:1) Fabrication and testing of a bi-enzyme electrochemical amplification module in a microfluidic format.2) Optimization of the various parameters affecting the performance of the amplification system.3) Demonstration of real-time detection of a model biological warfare simulant such as Bacillus globigii.There are two long-term goals after successful completion of this project. One is integration of automated sample collection utilizing MesoSystems' core technology in aerosol collection and particle concentration. The second is multiplexing of real-timedetection using a more sophisticated microfluidic cartridge. The incorporation of both of these advances will produce an inexpensive, handheld unit that can sample the air in any environment and perform rapid, sensitive detection of any biologicalparticles. The proposed amplification system will be capable of rapidly quantifying the presence of extremely low concentrations of a biological agent in a short amount of time. This product, the simplest version of a potential family of products, would itself bea highly marketable product. Areas of use include emergency response to a possible biological warfare threat and in microbiology laboratories. The system proposed here would be faster, cheaper, and more portable than most other detection schemes,including PCR.