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

这个小企业创新研究（SBIR）二期项目提议通过证明一种名为自交连锁反应（ACR）的创新技术的可行性，开发用于常规基因检测的新产品。ACR的智力优势在于它是一种无需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，并导致常规基因检测中基于分子的分析成本高，因为需要劳动密集型的样品制备和分析开发，以优化现有分子技术中的这些抑制剂。由于ACR不涉及任何对聚合酶抑制剂敏感的试剂，因此对样品制备的要求预计会很低，总体分析开发和测试周转时间预计会快得多。该项目的具体重点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扩增DNA靶序列。研究将使用低拷贝数目标核酸序列，其中包含跨多个位点的生物相关snp。预期的技术结果应显示在缺乏酶或核苷酸的情况下，对多个位点上的多个snp进行稳健、特异性和可重复的扩增。建议的活动有何更广泛的影响？如果成功，该项目的更广泛影响/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分析优化时间，显著提高效率和质量，降低临床诊断和常规基因检测的成本。由于非酶扩增加上ACR固有的简单性，使得该技术比现有的分子技术更适合于临床和实验室环境中不同样品类型的标准化，预计ACR技术将推动新一代分子诊断和筛选产品的发展。这些产品有望提供更有效、更简单、更便宜、更快速、更准确的常规基因检测，用于广泛的疾病和遗传性疾病的生物标志物，包括那些目前传统分子方法无法实现的疾病和遗传性疾病。因此，ACR不仅有可能在遗传水平上推进对疾病的理解，而且还将通过加强生物医学发现、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 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.