Sensitive detection of binding to DNA microarrays

DNA 微阵列结合的灵敏检测

• 批准号: 6738718
• 负责人: HADLEY D SIKES
• 金额: $ 3.97万
• 依托单位: UNIVERSITY OF COLORADO
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-11-16 至 2005-11-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6738718
• 关键词: biotechnology; diagnostic tests; fluorescent dye /probe; influenzavirus A; microarray technology; nucleic acid hybridization; nucleic acid quantitation /detection; photochemistry; polymerization; portable biomedical equipment; postdoctoral investigator; rapid diagnosis; technology /technique development; ultraviolet radiation; virus genetics

DESCRIPTION (provided by applicant): The purpose of the proposed project is to employ a surface-initiated photopolymerization reaction for highly sensitive detection of hybridization to a DNA microarray. DNA microarrays have the potential for high impact in health-related applications, however, a number of challenges must be overcome. One of these, and the focus of this proposal, is the development of a rapid, inexpensive and reliable method to detect a small number of hybridization events (approximately 100) for use in robust, portable, user-friendly devices. The proposed photopolymerization detection scheme will be tested in the Flu Chip, a novel inexpensive device designed to diagnose and subtype the influenza virus rapidly. Photoinitiators will be attached to the surface of DNA microarrays only in places where hybridization between the viral DNA of interest and the probe DNA of the microarray has occurred. Fluorescent monomer will be placed in contact with these hybridized arrays, and in the presence of ultraviolet radiation, a fluorescent polymer will grow from the surfacebound initiators. Unreacted monomers can be easily rinsed from areas of the array where no hybridization occurred, and the remaining surface-immobilized fluorescent polymer reveals which areas of the microarray contain captured viral genetic material.

描述（由申请人提供）：本项目的目的是采用表面引发的光聚合反应来高灵敏度地检测 DNA 微阵列的杂交。 DNA 微阵列在健康相关应用中具有巨大影响力的潜力，但必须克服许多挑战。其中之一，也是本提案的重点，是开发一种快速、廉价且可靠的方法来检测少量杂交事件（大约 100 个），以用于稳健、便携式、用户友好的设备。所提出的光聚合检测方案将在流感芯片中进行测试，这是一种新型廉价设备，旨在快速诊断流感病毒并对其进行分型。仅在感兴趣的病毒DNA与微阵列的探针DNA之间发生杂交的地方，光引发剂才会附着在DNA微阵列的表面上。荧光单体将与这些杂化阵列接触，并且在存在紫外线辐射的情况下，荧光聚合物将从表面结合的引发剂生长。未反应的单体可以很容易地从未发生杂交的阵列区域中冲洗掉，剩余的表面固定荧光聚合物揭示了微阵列的哪些区域含有捕获的病毒遗传物质。