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Switch on or switch off: An optical DNA sensor based on poly(p-phenylenevinylene) grafted magnetic beads

机译:开启或关闭:基于聚对苯乙撑乙烯接枝磁珠的光学DNA传感器

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摘要

There has been an enormous demand for commercial label-free DNA sensors in a diverse range of fields including pre-emptive medicine, diagnostics, environmental monitoring, and food industry. Addressing the need for sensitive, selective and facile DNA sensors, we demonstrate a novel switch on/off sensor design that utilizes sandwich hybridization between photoluminescent anionic conjugated polyelectrolyte (CPE) bound captureprobe coated onto magnetic beads, target and the signaling probe. The hybridization-readout in our sensor was monitored by either fluorescence resonance energy transfer (FRET, switch-on) or superquenching (switch-off) depending on the type of signaling probe used. Moreover recent designs that utilize beads for sensing DNA have been limited towards using electrostatic interactions or intercalation of dyes to observe FRET. To our knowledge this is the first report of a switch on/off sensor utilizing either FRET or superquenching thus providing flexibility for future development of such rapid, facile and sensitive DNA sensors. The FRET-based sensor was investigated by optimizing the reaction parameters and selectivity. A low detection limit of 240. fmol in 2. mL of SSC buffer was achieved.
机译:在包括先发制人的医学,诊断,环境监测和食品工业在内的各个领域,对无标记的商业化DNA传感器有巨大的需求。为了满足对灵敏,选择性和便捷的DNA传感器的需求,我们展示了一种新颖的开关传感器设计,该设计利用涂覆在磁珠,靶标和信号探针上的光致发光阴离子共轭聚电解质(CPE)结合捕获探针之间的夹心杂交。根据所用信号探针的类型,通过荧光共振能量转移(FRET,接通)或超猝灭(关断)来监测传感器中的杂交读数。此外,利用珠粒来感测DNA的最新设计已经限于使用静电相互作用或染料的嵌入来观察FRET。就我们所知,这是首次使用FRET或超淬火技术的开/关传感器的报告,从而为此类快速,便捷和灵敏的DNA传感器的未来开发提供了灵活性。通过优化反应参数和选择性研究了基于FRET的传感器。在2 mL SSC缓冲液中实现了240. fmol的低检测限。

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