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

这个小型企业创新研究(SBIR)第二阶段项目计划通过展示一种名为自动寡核苷酸链式反应(ACR)的创新技术的可行性，来开发用于常规基因测试的新产品。ACR的智力优势是一种不需要核苷酸或酶的核酸扩增技术。在许多实验室样本和临床样本中都发现了聚合酶抑制剂，这导致了常规基因测试中基于分子的分析的高成本，因为需要劳动密集型的样品制备和分析开发来利用当前的分子技术来优化这些抑制剂。由于ACR不涉及任何对聚合酶抑制剂敏感的试剂，对样品制备的要求预计较低，总体分析开发和测试周转时间预计会快得多。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，以及证明ACR可以在没有酶或核苷酸的情况下指数级扩增DNA靶序列。将使用低拷贝数的目标核酸序列进行研究，这些序列包含跨多个基因座的生物相关SNPs。预期的技术结果应该显示，在没有酶或核苷酸的情况下，多个基因座上的多个SNPs具有强大的、特异的和可重复性的扩增。拟议的活动有哪些更广泛的影响？如果该项目成功，其更广泛的影响/商业潜力是一种使能技术的创新，可以显著减少样品准备和分析优化时间，显著提高效率和质量，并降低临床诊断和常规基因测试的成本。由于非酶扩增加上ACR固有的简单性使这项技术比现有的分子技术更易于在不同样本类型的临床和实验室环境中标准化，因此预计ACR技术将推动新一代分子诊断和筛查产品的开发。预计这些产品将为各种疾病和遗传障碍提供更有效、更简单、更便宜、更快和更准确的常规基因检测，包括那些目前无法用传统分子方法获得的疾病和遗传障碍的生物标志物。因此，ACR不仅可能在基因水平上促进对疾病的理解，还将通过增强生物医学发现、诊断和个性化医学为人类健康和整个社会带来更广泛的好处。

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.