Sensitive detection of binding to DNA microarrays

描述(申请人提供)：该项目的目的是利用表面引发的光聚合反应来高灵敏地检测与DNA微阵列的杂交。DNA微阵列有可能在与健康相关的应用中产生很大的影响，然而，必须克服一些挑战。其中之一，也是这项建议的重点，是开发一种快速、廉价和可靠的方法来检测少量(约100个)杂交事件，用于坚固、便携、用户友好的设备。提出的光聚合检测方案将在流感芯片中进行测试，这是一种新型的廉价设备，旨在快速诊断流感病毒并进行亚型划分。光引发剂只有在感兴趣的病毒DNA与微阵列的探针DNA之间发生杂交的地方才会附着在DNA微阵列的表面。荧光单体将与这些杂化阵列接触，在紫外线照射下，荧光聚合物将从表面结合的引发剂中生长出来。未反应的单体可以很容易地从没有发生杂交的阵列区域中清洗出来，剩余的表面固定的荧光聚合物揭示了微阵列的哪些区域包含捕获的病毒遗传物质。

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.