A Nanomembrane-Based Nucleic Acid Sensing Platform

1065652 ChangIntellectual优点：便携式和多靶DNA/RNA诊断需要快速、现场可用、操作简单、可再生且经济的生物传感器系统。所提出的新平台是基于纳米多孔膜中几种新发现的电动现象的集成，这些现象有望将传统的无标记电化学和电容/电导生物传感器扩展到所需的鲁棒性、灵敏度和稳定性。（pM检测限约为一百万个分子），选择性（对于kb长的DNA/RNA，在约30个碱基的对接序列处仅具有一个错配的SNP辨别）和用于现场核酸检测的测定速度（5分钟）。这些新的检测功能的结果，非线性/选择性离子电导，动态控制离子耗尽/选择性，浓度极化，分子介电电泳，芯片上的pH值控制，电流体力学，表面电荷反转的核酸杂交所有新的电动和静电现象的纳米多孔膜，最近才被理解或发现的最新的纳米纤维和成像能力。拟议的工作将优化和集成这些新的物理现象到一个自动化的膜传感平台的手持DNA/RNA设备适合现场应用的科学仔细检查详细的非平衡电动现象，并通过利用最新的纳米/微加工技术。更广泛的影响：拟议的工作将提供博士和博士后学生与一个异常丰富的教育经验。它涉及新的电动力学和膜物理/化学的基础科学研究，最新的微/纳米制造技术，传染病健康科学，分子遗传学和基因组学，生态学和微型仪器设计，以开发可以对生物研究产生重大影响的原型，更商业化的是，生物技术工业部门。PI在主要研究型大学的终身教职（过去5年中有11名）中有着显著的记录，其中包括3名NSF职业奖获得者，3名女性和1名非洲裔美国人。教育机会也延伸到本科生和当地高中学生/教师通过积极的夏季外展计划在PI的实验室。几乎所有来自PI实验室的本科研究人员都进入了顶级博士课程。由于PI与台湾、韩国、欧洲和中国的机构有着密切的联系，因此国际合作的机会很大，因为他是美国物理研究所杂志《Biomicrofluidics》的创始人和主编，该杂志具有很高的影响力。一个可行的便携式用于病毒检测、细菌检测等的（手持式）和无标记DNA/RNA检测平台将推动生物传感器行业的重大技术进步，因为它将通过消除耗时的PCR或逆转录PCR步骤以及荧光传感平台的昂贵/笨重/人员密集型光学检测器而从根本上改变用于医学、环境、农业和生物防御应用的病原体检测方法。稳健和便携式RNA检测技术尚未出现，因为几个突出的技术挑战-缓慢的测定时间（长于RNA降解时间），对样品碎片和化学成分的敏感性，以及昂贵/笨重的检测仪器。拟议的项目调查，与互补的基础和制造的努力，几个新的纳米多孔膜现象，承诺减轻这些障碍，并将它们集成到一个多路复用芯片平台，可能会导致一个新的分子传感产品。因此，该项目可以影响纳米科学和纳米生物技术。

1065652ChangIntellectual Merit: Portable and multi-target DNA/RNA diagnostics requires a rapid, field-usable, simple to operate, regenerable and economical biosensor system. The proposed new platform is based upon the integration of several newly discovered electrokinetic phenomena in nanoporous membranes that promise to extend traditional label-free electrochemical and capacitance/conductance biosensors to the requisite robustness, sensitivity (pM detection limit of about one million molecules), selectivity (SNP discrimination with only one mismatch at a ~30-base docking sequence for a kb-long DNA/RNA) and assay speed (5 minutes) for on-field nucleic acid detection. These new detection features are results of nonlinear/selective ionic conductance, dynamically controlled ion depletion/selectivity, concentration polarization, molecular dielectrophoresis, on-chip pH control, electrohydrodynamics, surface charge inversion by nucleic acid hybridization all new electrokinetic and electr static phenomena of nanoporous membranes that are only recently understood or discovered with the latest nanofabrication and imaging capabilities. The proposed work will optimize and integrate these new physical phenomena into an automated membrane sensing platform for hand-held DNA/RNA devices suitable for field applications by scientifically scrutinizing the detailed non-equilibrium electrokinetic phenomena and by exploiting the latest nano/microfabrication technologies.Broader Impacts: The proposed work will provide PhD and post-doc students with an unusually rich educational experience. It involves fundamental scientific studies of new electrokinetic and membrane physics/chemistry, the latest micro/nano-fabrication technologies, contagious disease health science,molecular genetics and genomics, ecology and miniature instrumentation design to develop prototypes that can have a significant impact on biological research and, more commercially, the biotechnology industry sector. The PI has significant track records of placing group members in tenure-track faculty positions (11 in the last 5 years) at major research universities, including 3 NSF Career Awardees, 3 women and 1 African American. Educational opportunities also extend to undergraduates and local high school students/teachers through an active summer outreach program in the PI's laboratory. Almost all of the undergraduate researchers from the PI's lab go on to top PhD programs. Significant opportunities exist for international collaborations due to the PI's strong ties to institutions in Taiwan, Korea, Europe, and China through his capacity as the founding and chief editor of Biomicrofluidics, an American Institute of Physics journal with high impact factor.Scientific and Technological Impact: A viable portable (handheld) and label-free DNA/RNA detection platform for viral assays, bacteria detection etc. will spur a major technological advance in the biosensor industry, as it will fundamentally transform pathogen detection methodology for medical, environmental, agricultural and biodefense applications by eliminating the time-consuming PCR or reverse transcription PCR step and the costly/bulky/personnel-intensive optical detectors of fluorescent sensing platforms. Robust and portable RNA detection technologies have yet to appear because of several outstanding technological challenges---slow assay time (longer than RNA degradation time), sensitivity to sample debris and chemical content, and expensive/bulky detection instrumentation. The proposed project investigates, with complementary fundamental and fabrication efforts, several new nanoporous membrane phenomena that promise to alleviate these obstacles and integrates them onto a multiplex chip platform that may lead to a new molecular sensing product. The project can hence impact both nano science and nano biotechnology.