Département de Pharmacochimie Moléculaire UMR 5063, Centre National de la Recherche Scientifique, University Grenoble Alpes Grenoble, France.
Laboratoire ARNA, Institut National de la Santé et de la Recherche Médicale U869, Université Bordeaux Bordeaux, France.
Front Chem. 2015 Aug 12;3:49. doi: 10.3389/fchem.2015.00049. eCollection 2015.
Here, we report a new homogeneous DNA amplification-based aptamer assay for small analyte sensing. The aptamer of adenosine chosen as the model analyte was split into two fragments able to assemble in the presence of target. Primers were introduced at extremities of one fragment in order to generate the amplifiable DNA component. The amount of amplifiable fragment was quantifiable by Real-Time Polymerase Chain Reaction (RT-PCR) amplification and directly reliable on adenosine concentration. This approach combines the very high separation efficiency and the homogeneous format (without immobilization) of capillary electrophoresis (CE) and the sensitivity of real time PCR amplification. An ultrafast isolation of target-bound split aptamer (60 s) was developed by designing a CE input/ouput scheme. Such method was successfully applied to the determination of adenosine with a LOD of 1 μM.
在这里，我们报告了一种新的基于均相 DNA 扩增的适体分析方法，用于小分子分析物的传感。所选的腺苷适体被分割成两个片段，能够在目标存在的情况下组装。引物被引入一个片段的末端，以便产生可扩增的 DNA 成分。可扩增片段的量可以通过实时聚合酶链反应（RT-PCR）扩增来定量，并直接依赖于腺苷浓度。这种方法结合了毛细管电泳（CE）的高分离效率和均相格式（无需固定化）以及实时 PCR 扩增的灵敏度。通过设计 CE 输入/输出方案，开发了一种超快速的目标结合分裂适体的分离方法（60 秒）。该方法成功地应用于腺苷的测定，LOD 为 1 μM。
