Castañeda María Teresa, Alegret Salvador, Merkoçi Arben
Nanobioelectronics & Biosensors Group, Institut Catalá de Nanotecnologia, Barcelona, Catalonia, Spain.
Methods Mol Biol. 2009;504:127-43. doi: 10.1007/978-1-60327-569-9_9.
A novel, rapid, and sensitive protocol for the electrochemical detection of DNA hybridization that take the advantage of a magnetic separation/mixing process and the use of monomaleimido-gold nanoparticles of 1.4 nm diameter as label is presented. A sandwich-type assay is formed in this protocol by the capture probe DNA immobilized on the surface of magnetic beads and the double hybridization of the target (cystic fibrosis related DNA), first with the immobilized probe, and then with signaling probe DNA labeled with monomaleimido-gold nanoparticles. When the assay is completed, the final conjugate is transferred onto genomagnetic sensor surface (graphite epoxy composite electrode with a magnet inside) used as working electrode, and then the direct determination of gold nanoparticles by differential pulse voltammetry striping technique is carried out. This protocol is quite promising for numerous applications in different fields as clinical analysis, environmental control as well as other applications.
本文提出了一种新型、快速且灵敏的电化学检测DNA杂交的方法,该方法利用了磁分离/混合过程,并使用直径为1.4 nm的单马来酰亚胺金纳米颗粒作为标记物。在此方法中,通过固定在磁珠表面的捕获探针DNA以及靶标(囊性纤维化相关DNA)的双重杂交形成夹心型检测法,靶标首先与固定的探针杂交,然后与用单马来酰亚胺金纳米颗粒标记的信号探针DNA杂交。检测完成后,将最终的复合物转移到用作工作电极的基因组磁传感器表面(内部带有磁体的石墨环氧复合电极),然后通过差分脉冲伏安法溶出技术直接测定金纳米颗粒。该方法在临床分析、环境监测等不同领域的众多应用中颇具前景。