Technology Development

项目摘要：技术发展这一更新申请的重点是开发能够以大大降低的成本研究人类基因组和人类疾病潜在机制的技术。分析人体样品的方法必须非常灵敏，以便：1)检测低丰度的生物分子；2)使用小样本量提供信息。然而，目前的基因组学和蛋白质组学方法在分析人类样本的适用性方面差异很大。例如，基因组方法如微阵列和下一代测序具有高灵敏度，但由于产生可检测信号所需的DNA扩增策略的偏差，其定量价值受到限制。相比之下，目前的蛋白质组学方法，如酶联免疫吸附试验，完全缺乏灵敏度，使得可能作为信息性生物标志物的低丰度蛋白质未被检测到。在提案的第二层。技术开发，我们专注于开发能够检测核酸和蛋白质的技术，不仅具有较高的准确性和灵敏度，而且需要最少的时间和费用，以促进其转化为临床医学。这些技术将为生物医学研究人员提供更好的工具来调查人类疾病的过程，并使他们能够使用侵入性较小的诊断材料来源，如汗液、唾液或呼吸。这些技术在之前的资助期间处于创新阶段，并达到了预期的里程碑。目前的发展方向将是进一步加强这些工具，并证明它们比现有技术有所改进。我们将开发一种基于改进原子力显微镜的方法，用于核酸的无标记纳米力学定量，以及两种集成微流体和电阻抗传感的方法，用于高通量数字检测微小样品中的蛋白质。一旦建立了原理证明，这些技术将准备出口并应用于特定的临床问题。

Project Summary: Technology Development The focus of this renewal application is on the development of technologies that will enable the study of the human genome and the mechanisms underlying human disease at a substantially-reduced cost. Methods to analyze human samples must be extremely sensitive in order to: 1) detect low abundance biomolecules; and 2) be informative using small sample volumes. However, current genomic and proteomic methods vary widely in their suitability for analyzing human samples. For example, genomic methods such as microarrays and next generation sequencing have high sensitivity, but are limited in their quantitative value because ofthe biases of DNA amplification strategies that are required to generate detectable signals. In contrast, current proteomic methods such as ELISA assays simply lack sensitivity altogether, leaving the low abundance proteins that may be informative biomarkers undetected. In this second tier of the proposal. Technology Development, we are focused on developing technologies that can detect nucleic acids and proteins not only with high accuracy and sensitivity, but also with a minimum of time and expense in order to facilitate their translation to clinical medicine. Such technologies will equip biomedical researchers with better tools to investigate the human disease process, and enable the use of less invasive sources of diagnostic material, such as sweat, saliva, or breath. These technologies were in the innovation phase during the previous funding period and met desired milestones. Current development will be directed toward further strengthening the tools and demonstrating their improvement over existing technologies. We will develop a method based on modified atomic force microscopy for the label-free nanomechanical quantitation of nucleic acids, and two methods that integrate microfluidics and electrical impedance sensing for the high-throughput digital detection of proteins from minute samples. Once proof-of-principle has been established, these technologies will be primed for export and application to specific clinical problems.