BioNano Research Center, Korea Research Institute of Bioscience and Biotechnology, 111 Gwanhangno, Yuseong-gu, Daejeon 305-806, South Korea.
Biosens Bioelectron. 2011 Jan 15;26(5):2125-9. doi: 10.1016/j.bios.2010.09.018. Epub 2010 Sep 16.
For previously reported fluorescence Pb(2+) sensors, DNAzymes have lead to a significant increase in Pb(2+) detecting sensitivity and specificity. However, these sensors suffer from incomplete fluorescence quenching and require additional steps for annealing DNAzymes and substrates as well as for removing the uncoupled substrates. In this study, we successfully overcome these issues by immobilizing the substrate nucleic acids on gold nanoparticles through thiol linkages. The immobilization of the substrate molecules to the gold nanoparticles lead to almost-complete fluorescence quenching and fast Pb(2+) detection, without altering the Pb(2+) specificity of the DNAzymes. After optimizing the concentration of DNAzymes, reaction time and pH, we could detect Pb(2+) as low as 5 nM within 20 min without the preliminary and the post treatments. Considering the multi-color-fluorescence quenching capability of gold nanoparticles and the to-be-developed functional nucleic acids for other metal ions, this study could extend the application of DNAzymes to the detection of multiple heavy metal ions.
对于之前报道的荧光 Pb(2+)传感器,DNA 酶显著提高了 Pb(2+)检测的灵敏度和特异性。然而,这些传感器存在荧光猝灭不完全的问题,并且需要额外的步骤来退火 DNA 酶和底物,以及去除未偶联的底物。在这项研究中,我们通过将底物核酸通过巯基键合固定在金纳米粒子上,成功地克服了这些问题。底物分子固定在金纳米粒子上导致几乎完全的荧光猝灭和快速的 Pb(2+)检测,而不改变 DNA 酶对 Pb(2+)的特异性。在优化 DNA 酶的浓度、反应时间和 pH 值后,我们可以在 20 分钟内检测到低至 5 nM 的 Pb(2+),而无需进行预处理和后处理。考虑到金纳米粒子的多色荧光猝灭能力以及正在开发的用于其他金属离子的功能核酸,这项研究可以将 DNA 酶的应用扩展到多种重金属离子的检测。