High Throughput Isothermal Monitoring

DESCRIPTION (provided by applicant): The long-term goal of this work is to develop a high-throughput device for the rapid and specific detection of viral or bacterial loads from blood or tissue samples, with the use of PCR or other target amplification. The RiboMaker (tm) Pathogen Detection System, RiboPATH (tm), currently under development at Designer Genes, Inc., is a rapid, isothermal, signal amplification system for the detection of target DNA or RNA that should be the ideal signal generation chemistry for this device. It is not a target amplification technology, nor a true signal amplification technology. RiboPATH (tm) utilizes a novel and proprietary process to create thousands to millions of detectable signals for each target. These signals are generated by short oligonucleotides that are reiteratively synthesized and released by our novel RiboMaker (tm) polymerase when linked to the molecular target. In the RiboPATH (tm) Fluorescent Resonance Energy Transfer (FRET) system, the oligonucleotides produced are tagged at both the 5' and 3' ends with a set of fluorescent dyes that serve as an efficient fluorescent energy donor/acceptor pair. The dyes are enzymatically incorporated into the signal-generating oligonucleotides in a target-dependent process. Only when the two fluorescent nucleotide analogs are joined together covalently will the donor and acceptor pair come close enough together for energy transfer (FRET). Signals from these oligonucleotide products can be measured in real time without removal of the unincorporated nucleotide analog substrates. The goal of this Phase 1 proposal is to optimize the first generation of the RiboPATH (tm) detection technology.