This Small Business Innovation Research (SBIR) Phase I project will provide an opportunity to develop novel products for routine genetic testing by demonstrating feasibility of an innovative biotechnology 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.
The broader impact/commercial potential of this project 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. Non-enzymatic amplification coupled with the inherent simplicity of ACR makes this technology more amenable to standardizing in clinical and lab settings across different sample types as compared with existing molecular technologies. It is expected that ACR technology will drive the development of a new generation of molecular diagnostic and screening products towards more efficient, simpler, cheaper, faster, and more accurate routine genetic testing. The technology will be applicable to 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 potentially will not only advance our 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.
The outcome of the Small Business Innovation Research (SBIR) Phase I project demonstrated feasibility of an innovative amplification technology. The intellectual merit of the technology is that no enzymes or single-nucleotides are required. High cost of enzyme-based polymerase chain reaction (PCR) technology makes routine biomarker screening of samples too expensive. Contributing factors are high assay reagent costs and the need to develop specific protocols to remove polymerase inhibitors from samples. They ultimately make PCR non-scalable to the needs of laboratories involved in routine genetic testing. The technology developed from this SBIR project will significantly reduce direct assay cost by omitting expensive reagents including enzymes, single nucleotides, probes, and intercalating agents from the reaction. It will also obviate the need for master mixes, long thermo-extension steps, and protocols to remove PCR polymerase inhibitors from many sample types. Therefore, this technology overcomes sample-preparation bottlenecks, simplifies liquid handling steps, and shortens reaction cycles. The broader impact/commercial potential of this project is the innovation of a universal assay technology that will significantly reduce per-data-point and upfront assay optimization costs, shorten time-to-answer, and increase specificity in biomarker screening. The outcome is enabling products to exponentially increase genetic biomarker screening in routine settings with fewer resources. As a result, the technology will potentially not only advance our 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.
