SBIR Phase II: Autoligation Chain Reaction: DNA Amplification Without Enzymes or Nucleotides

This Small Business Innovation Research (SBIR) Phase II project proposes to develop novel products for routine genetic testing by demonstrating feasibility of an innovative technology called Autoligation Chain Reaction (ACR). The intellectual merit of ACR is an enabling nucleic acid amplification technology that requires no nucleotides or enzymes. Polymerase inhibitors are found in many laboratory samples and clinical specimens, and contribute to the high cost of molecular-based assays in routine genetic tests because labor-intensive sample preparation and assay development are required to optimize around these inhibitors with current molecular technologies. Because ACR does not involve any reagents sensitive to polymerase inhibitors, the requirement for sample preparation is expected to be low and overall assay development and testing turnaround times are expected to be much faster. Specific key research objectives of the project include design and synthesis of thermal-stable ACR reagents, and the demonstration that ACR can exponentially amplify DNA target sequences without enzymes or nucleotides. Research will be carried out using low copy-number target nucleic acid sequences containing bio-relevant SNPs across multiple loci. The anticipated technical results should show robust, specific, and reproducible amplifications of multiple SNPs on multiple loci in the absence of enzymes or nucleotides. What are the broader impacts of the proposed activity? The broader impact/commercial potential of this project, if successful, is the innovation of an enabling technology that could dramatically reduce sample preparation and assay optimization times, and significantly increase the efficiency and quality, and lower the cost of clinical diagnostics and routine genetic testing. Since non-enzymatic amplification coupled with the inherent simplicity of ACR makes this technology more amenable to standardization in clinical and lab settings across different sample types than existing molecular technologies, it is expected that ACR technology will drive the development of a new generation of molecular diagnostic and screening products. These products are expected to provide more efficient, simpler, cheaper, faster, and accurate routine genetic testing of a broad range of biomarkers for a wide range of diseases and genetic disorders, including those currently unattainable by traditional molecular methods. As a result, ACR will potentially not only advance the understanding of diseases at the genetic level, but also bring broader benefits to human health and society at large through enhanced biomedical discovery, diagnostics, and personalized medicine.