High throughput nucleic acid detection systems based on multiplexing using quantum dots

基于使用量子点的多重检测的高通量核酸检测系统

• 批准号: 336239-2006
• 负责人: Krull, Ulrich
• 金额: $ 7.43万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Strategic Projects - Group
• 财政年份: 2007
• 资助国家: 加拿大
• 起止时间: 2007-01-01 至 2008-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=363615
• 关键词: throughput; nucleic; acid; detection; systems

Nucleic acid sequences can be used for detection and identification of bacteria and viruses, and identificationof species and species modifications/mutations. Existing technologies (e.g. microarrays, multiplexedbead-based assays, and single use solution assays) are not ideally suited for the demands of many areas ofapplication. It is the combination of cost-effective, repetitive, fast, selective and quantitative response for lowquantities of nucleic acid sequences that is beyond reach of conventional methods, yet which is critical foranalyses in many areas of food safety and quality, clinical diagnostics and biosafety. The goal of the proposedresearch is to develop a quantitative high-throughput nucleic acid diagnostic system that is suitable for rapiddetection of well-defined target nucleic acids. Fluorescent semiconductor quantum dots (QD) will be coatedwith fluorescent nucleic acid probe molecules to create nanoscale biosensors that operate by emitting certaincolours of light when the probe molecules bind to target nucleic acid sequences (hybridization). The QDs willserve as excitation sources to stimulate emission from intrinsically labelled DNA probes when hybridizationoccurs, so that labelling of target nucleic acids will not be necessary. Excitation will be based on FluorescenceResonance Energy Transfer (FRET) from the QDs to the labels that are tethered to the probe molecules. QDs ofdifferent size offer different narrow band emission wavelengths using only one excitation wavelength. Uniquelabelled DNA probe sequences will be associated with differently sized QDs to prepare distinctive QDbiosensors. Mixtures of such QD biosensors have multi-colour emission and a number of target nucleic acidscan be simultaneously determined. Ensembles of such mixtures of different QD biosensors will beimmobilized within microfluidics channels. Electroosmotically driven fluid flow will be used to achievedynamic manipulation of stringency to control selectivity and regeneration of the QD biosensors. The intentionis to create a reversible high throughput sensing system that is suitable for concurrent analysis of multiplenucleic acid targets without the need for labelling of the targets.

核酸序列可用于细菌和病毒的检测和鉴定，以及物种和物种修饰/突变的鉴定。现有的技术（如微阵列、基于多功能微珠的检测和一次性溶液检测）并不理想地适合许多应用领域的需求。它结合了传统方法所无法达到的低成本、重复性、快速、选择性和定量的核酸序列分析，在食品安全和质量、临床诊断和生物安全的许多领域都是至关重要的。本研究的目标是建立一种高通量的核酸定量诊断系统，用于快速检测目标核酸。荧光半导体量子点（QD）将被涂上荧光核酸探针分子，以创建纳米级生物传感器，当探针分子与靶核酸序列结合（杂交）时，该传感器通过发射某些颜色的光来工作。当杂交发生时，量子点将作为激发源，刺激内在标记的DNA探针发射，因此不需要标记靶核酸。激发将基于从QD到拴系到探针分子的标记的共振能量转移（FRET）。不同尺寸的量子点在同一激发波长下可以提供不同的窄带发射波长。独特的DNA探针序列将与不同大小的量子点相关联，以制备独特的量子点生物传感器。这种量子点生物传感器的混合物具有多色发射，并且可以同时测定许多靶核酸。不同QD生物传感器的这种混合物的集合将被固定在微流体通道内。电驱动的流体流动将被用于严格性的电化学操纵，以控制QD生物传感器的选择性和再生。本发明的目的是创造一种可逆的高通量传感系统，其适合于同时分析多个核酸靶，而不需要标记靶。