Rapid, Binary-Mode Nucleic Acid Sequence Detection

快速二元模式核酸序列检测

• 批准号: 1265061
• 负责人: Harold Monbouquette
• 金额: $ 34.05万
• 依托单位: University of California-Los Angeles
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-01 至 2016-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1265061&HistoricalAwards=false
• 关键词: Rapid; Binary; Mode; Nucleic; Acid

Technical abstractDetection of specific nucleic acid (NA) sequences is crucial to infectious diseasediagnostics, genetic screening for hereditary diseases, personalized medicine, criminology,paternity disputes, and animal and plant breeding, among many other applications. Althoughpolymerase chain reaction (PCR) technology, coupled with oligonucleotide array chips, hasenabled specific identification of multiple sequences present in a sample at very low copynumber, this method requires added labels and reagents to detect the target NA. These labelsand the hardware (e.g., optics) required to sense them significantly increase the analysis time,cost, and footprint of devices for detecting specific NA sequences. As an alternative to current approaches, this project will develop novel nanopores capable of detecting pathogenic nucleic acids with single molecule resolution. These nanopores will be fabricated in silicon and will be engineered to contain beads labeled with complementary NA sequences. Binding of target oligonucleotides to the beads will result in the change of electrophoretic properties and will lead to nanopores becoming occluded by the beads. This event will be detected electrochemically for binary yes/no readout of the target NA. Non-technical abstractThere is a need to develop simple and inexpensive methods for detecting pathogens for medical or food safety applications. This proposal will develop a new approach based on small pores containing beads. The beads will react to pathogen biomarkers, will clog the pores and will result in the change of electrical current. This method is envisioned to have improved sensitivity and be simpler than the current approaches.

技术摘要特定核酸序列的检测对于传染病诊断、遗传性疾病的基因筛查、个性化医疗、犯罪学、亲子关系纠纷、动植物育种以及许多其他应用至关重要。尽管聚合酶链反应（PCR）技术与寡核苷酸芯片相结合，能够特异性地鉴定一个样本中存在的多个拷贝数很低的序列，但这种方法需要添加标记和试剂来检测目标NA。这些标签和硬件（例如，光学器件）显著增加了用于检测特定NA序列的装置的分析时间、成本和占地面积。 作为当前方法的替代方案，该项目将开发能够以单分子分辨率检测致病核酸的新型纳米孔。 这些纳米孔将在硅中制造，并将被工程化以包含用互补NA序列标记的珠。 靶寡核苷酸与珠的结合将导致电泳性质的变化，并将导致纳米孔被珠堵塞。 将通过电化学检测该事件，以获得目标NA的二进制是/否读数。 非技术摘要有必要开发简单和廉价的方法来检测病原体的医疗或食品安全应用。 该提案将开发一种基于含有珠粒的小孔的新方法。 珠子将与病原体生物标志物反应，将堵塞孔并将导致电流的变化。 该方法被设想为具有改进的灵敏度并且比当前的方法更简单。