Visual detection of labeled oligonucleotides using visible-light-polymerization-based amplification
Visual detection of labeled oligonucleotides using visible-light-polymerization-based amplification
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DOI：
10.1021/bm700672z
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发表时间：
2008-01-01
影响因子：
6.2
通讯作者：
Bowman, Christopher N.
中科院分区：
文献类型：
--
作者：
Hansen, Ryan R.;Sikes, Hadley D.;Bowman, Christopher N.
DNA生物芯片技术具有高度平行、快速和灵敏的靶病原体和疾病生物标志物的遗传诊断筛选的潜力。主要限制涉及使用仅实施廉价检测仪器的临床上适当的检测方法同时序列特异性鉴定低拷贝数靶多核苷酸。在这里，一个快速（20分钟），非酶的信号放大方法，利用表面引发的光聚合是在玻璃微阵列正式。使用共价偶联至链霉亲和素的可见光光引发剂来结合生物素标记的捕获序列。通过随后与单体溶液接触并适当曝光以仅从含有生物素标记的DNA的点产生20-240 nm厚的水凝胶层来实现扩增。观察到10（6）至10（7）的扩增因子，以及使用最小仪器（如光学显微镜或CCD相机）从低至类似于10（4）的标记寡核苷酸产生的可检测响应。
DNA biochip technology holds potential for highly parallel, rapid, and sensitive genetic diagnostic screening of target pathogens and disease biomarkers. A primary limitation involves a simultaneous, sequence-specific identification of low copy number target polynucleotides using a clinically appropriate detection methodology that implements only inexpensive detection instrumentation. Here, a rapid (20 min), nonenzymatic method of signal amplification utilizing surface-initiated photopolymerization is presented in glass microarray formal. Visible light photoinitiators covalently coupled to streptavidin were used to bind biotin-labeled capture sequences. Amplification was achieved through subsequent contact with a monomer solution and the appropriate light exposure to generate 20-240-nm-thick hydrogel layers exclusively from spots containing the biotin-labeled DNA. An amplification factor of 10(6) to 10(7) was observed as well as a detectable response generated from as low as similar to 10(4) labeled oligonucleotides using minimal instrumentation, such as an optical microscope or CCD camera.