PIEZO ELECTRIC PIPETTING TECHNOLOGY FOR DNA ANALYSIS

用于 DNA 分析的压电移液技术

• 批准号: 6015579
• 负责人: PETER J. WIKTOR
• 金额: $ 13.09万
• 依托单位: ENGINEERING ARTS
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-01 至 2001-04-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6015579
• 关键词: DNA; biomedical automation; biomedical equipment development; computer program /software; computer system design /evaluation; computer system hardware; fluid flow; gene expression; genetic polymorphism; nucleic acid hybridization; nucleic acid quantitation /detection; polymerase chain reaction; technology /technique development

This grant brings together two innovative technologies: a novel fluid transfer device and a novel technique for detecting Single Nucleotide Polymorphisms (SNPs) on cDNA arrays. Integrated together, these two technologies will have an immediate and significant impact on the molecular analysis of cancer. Actuated by piezo-electric elements, the fluid transfer technology enables the pipetting of sub-nanoliter volumes of fluid. Additionally, novel sensing technology allows the operational state of the device to be continuously monitored. The sensing technology is indispensable for massively parallel DNA analysis applications requiring thousands of different samples to be reliably aspirated and dispensed. The ultimate goal is to integrate the fluid transfer and sensor technology into an automated pipetting instrument for cancer research and general laboratory applications. Initially the instrument will be applied to a novel technology making it possible for the first time to detect SNPs, deletions and insertions on readily available and relatively inexpensive, spotted cDNA arrays. An Invention Disclosure has been filed by the Collaborating Investigator with the Fred Hutchinson Cancer Research Center (F.H.C.R.C.), Seattle, WA for this exciting new technology. The detection of SNPs on DNA microarrays has heretofore only been possible using allele specific hybridization on oligonucleotide arrays (GeneChips) produced by Affymetrix. Limited access to these GeneChips and their prohibitive costs will make it difficult to apply this technology to large numbers of DNA samples. A major goal of the proposed study is to develop and demonstrate the feasibility of the new SNP detection methodology for high throughput detection of polymorphisms at multiple loci using the high-density cDNA arrays produced by the novel arrayer to be developed in this grant. These highly reproducible, high-density arrays are also expected to provide improved sensitivity and specificity in the analysis of mRNA expression. Experiments will be performed to demonstrate the advantages of the proposed technology over existing pin based mechanical arrayers for gene expression analysis. PROPOSED COMMERCIAL APPLICATIONS: Fueled by emerging technologies like bio-chips, micro-arrays and high throughput screening and the high costs of reagents in general, the bio- medical industry is going towards assays using smaller fluid sample volumes. The proposed research directly addresses this growing market by developing a general purpose automated laboratory pipetting instrument for these applications. By keeping development costs down, the new technology will make high quality, high density DNA arrays accessible to a much broader segment of the cancer research community.

这项资助汇集了两项创新技术：一种新型流体转移装置和一种用于检测 cDNA 阵列上的单核苷酸多态性 (SNP) 的新技术。这两种技术结合在一起将对癌症的分子分析产生直接而重大的影响。流体传输技术由压电元件驱动，能够移取亚纳升体积的流体。此外，新颖的传感技术可以持续监控设备的运行状态。对于需要可靠吸取和分配数千个不同样本的大规模并行 DNA 分析应用来说，传感技术是不可或缺的。最终目标是将流体传输和传感器技术集成到用于癌症研究和一般实验室应用的自动移液仪器中。最初，该仪器将应用于一项新技术，首次能够在现成且相对便宜的斑点 cDNA 阵列上检测 SNP、缺失和插入。合作研究人员已向华盛顿州西雅图 Fred Hutchinson 癌症研究中心 (F.H.C.R.C.) 提交了一项发明披露，以表彰这项令人兴奋的新技术。迄今为止，DNA 微阵列上的 SNP 检测只能使用 Affymetrix 生产的寡核苷酸阵列 (GeneChips) 上的等位基因特异性杂交来实现。这些基因芯片的获取途径有限且成本高昂，因此很难将该技术应用于大量 DNA 样本。拟议研究的一个主要目标是开发并证明新的 SNP 检测方法的可行性，该方法可使用本次资助中将开发的新型阵列仪产生的高密度 cDNA 阵列来高通量检测多个位点的多态性。这些高度可重复的高密度阵列也有望在 mRNA 表达分析中提供更高的灵敏度和特异性。将进行实验以证明所提出的技术相对于现有的基于针的机械阵列器在基因表达分析方面的优势。拟议的商业应用：在生物芯片、微阵列和高通量筛选等新兴技术以及一般试剂成本较高的推动下，生物医学行业正在转向使用更小液体样本量的检测。拟议的研究通过开发适用于这些应用的通用自动化实验室移液仪器来直接解决这个不断增长的市场。通过降低开发成本，新技术将使癌症研究界更广泛的领域能够获得高质量、高密度的 DNA 阵列。