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

这个小企业创新研究（SBIR）第一阶段项目将提供一个机会，通过展示称为自动连接链反应（ACR）的创新生物技术的可行性，开发用于常规基因检测的新产品。 ACR的智力价值是一种不需要核苷酸或酶的核酸扩增技术。 聚合酶抑制剂存在于许多实验室样品和临床标本中，并且导致常规遗传检测中基于分子的测定的高成本，因为需要劳动密集型样品制备和测定开发以利用当前分子技术围绕这些抑制剂进行优化。 由于ACR不涉及任何对聚合酶抑制剂敏感的试剂，因此预计对样品制备的要求较低，总体检测开发和检测周转时间预计将更快。 该项目的具体关键研究目标包括设计和合成热稳定的ACR试剂，并证明ACR可以在没有酶或核苷酸的情况下以指数方式扩增DNA靶序列。 研究将使用包含跨多个基因座的生物相关SNP的低拷贝数靶核酸序列进行。 预期的技术结果应显示在不存在酶或核苷酸的情况下多个基因座上的多个SNP的稳健、特异性和可再现的扩增。该项目更广泛的影响/商业潜力是一种使能技术的创新，可以大大减少样品制备和测定优化时间，显着提高效率和质量，降低临床诊断和常规基因检测的成本。 与现有的分子技术相比，非酶促扩增加上ACR的固有简单性使得该技术更适合在临床和实验室环境中跨不同样品类型进行标准化。 预计ACR技术将推动新一代分子诊断和筛查产品的发展，使其朝着更高效、更简单、更便宜、更快速、更准确的常规基因检测方向发展。 该技术将适用于广泛的疾病和遗传性疾病的广泛生物标志物，包括目前传统分子方法无法实现的生物标志物。 因此，ACR不仅有可能在遗传水平上促进我们对疾病的理解，而且还可以通过增强生物医学发现，诊断和个性化医疗为人类健康和整个社会带来更广泛的利益。

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.