MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, Guangzhou 510631, China.
Anal Chim Acta. 2012 Feb 3;713:45-9. doi: 10.1016/j.aca.2011.11.003. Epub 2011 Nov 25.
In this study, we firstly demonstrated that Bst DNA polymerase shows specific recognition and function on the T-Hg(2+)-T biomimetic structure. Based on this, a novel available electrochemiluminescence (ECL) sensor for Hg(2+) has been developed. In this strategy, magnet beads tagged primer was designed to complementary to the region of the circular padlock probe but with two T-T mismatches at the 3' end. The mismatched primers cannot be extended by Bst DNA polymerase in the absence of Hg(2+). Stable T-Hg(2+)-T can be formed in the presence of Hg(2+), thus induces the elongation and amplification reaction by DNA polymerase with a rolling circular amplification (RCA) mechanism. Subsequently, the resulted RCA products are hybridized with the tris (bipyridine) ruthenium (TBR)-tagged probes and detected by ECL platform. Current method shows a sub-nanomolar sensitivity and excellent selectivity over a spectrum of interference metal ions.
在这项研究中,我们首先证明了 Bst DNA 聚合酶对 T-Hg(2+)-T 仿生结构具有特异性识别和功能。基于此,我们开发了一种新型的电化学发光(ECL)传感器用于检测 Hg(2+)。在该策略中,设计了带有磁珠标记的引物使其与环型发夹探针的区域互补,但在 3'端有两个 T-T 错配。在没有 Hg(2+)的情况下,Bst DNA 聚合酶不能延伸错配引物。在存在 Hg(2+)的情况下,可以形成稳定的 T-Hg(2+)-T,从而通过 DNA 聚合酶诱导延伸和扩增反应,采用滚环扩增(RCA)机制。随后,所得的 RCA 产物与三联吡啶钌(TBR)标记的探针杂交,并通过 ECL 平台进行检测。该方法对一系列干扰金属离子具有亚纳摩尔级的灵敏度和优异的选择性。