Polymer Electronics Research Centre, School of Chemical Sciences, The University of Auckland, Private Bag 92019, Auckland, New Zealand.
Polymer Electronics Research Centre, School of Chemical Sciences, The University of Auckland, Private Bag 92019, Auckland, New Zealand; Key Laboratory of Polarized Materials and Devices, Ministry of Education, East China Normal University, Shanghai 200062, China.
Biosens Bioelectron. 2012 May 15;35(1):498-502. doi: 10.1016/j.bios.2012.03.022. Epub 2012 Mar 20.
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.
在包括预防性医疗、诊断、环境监测和食品工业在内的众多领域,人们对商用无标记 DNA 传感器有着巨大的需求。为满足对敏感、选择性和简便 DNA 传感器的需求,我们展示了一种新颖的开启/关闭传感器设计,该设计利用光致阴离子共轭聚合物(CPE)与结合在磁珠上的捕获探针、目标物和信号探针之间的三明治杂交。我们的传感器中的杂交读出通过荧光共振能量转移(FRET,开启)或超猝灭(关闭)来监测,具体取决于所使用的信号探针的类型。此外,最近利用珠子进行 DNA 感测的设计仅限于利用静电相互作用或染料的嵌入来观察 FRET。据我们所知,这是第一个利用 FRET 或超猝灭的开启/关闭传感器的报告,从而为未来开发这种快速、简便和灵敏的 DNA 传感器提供了灵活性。通过优化反应参数和选择性,对基于 FRET 的传感器进行了研究。在 2mL SSC 缓冲液中实现了 240fmol 的低检测限。