Label-free impedance detection of oligonucleotide hybridisation on interdigitated ultramicroelectrodes using electrochemical redox probes

被引:62
作者
Dharuman, V
Grunwald, I
Nebling, E
Albers, J
Blohm, L
Hintsche, R
机构
[1] Fraunhofer Inst Silicon Technol, Dept Biotech Microsyst, BTMS, D-25524 Itzehoe, Germany
[2] eBiochip Syst GmbH, D-25524 Itzehoe, Germany
关键词
ODN hybridisation; label-free detection; IDA electrodes; Si-technology; impedance; electrical biochips;
D O I
10.1016/j.bios.2004.12.020
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
The direct detection of oligodeoxynucleotide (ODN) hybridisation using electrochemical impedance spectroscopy was made on interdigitated array (IDA) gold (An) ultramicroelectrodes manufactured by silicon technology. The immobilisation of single stranded ODNs (ssODNs) was accomplished by self-assembling of thiol-modified ODNs onto an Au-electrode surface. Faradaic impedance was measured in the presence of K-3 [Fe(CN)(6)]. Double strand formation was identified by a decrease of approximately 50% in impedance in the low frequency region in the presence of K-3[Fe(CN)(6)], compared to the spectrum of single stranded ODN. The frequency dependent diffusion of Fe(CN)(6)(3-) ions through defects in the ODN monolayer determines the impedance of Au-ssODN surface. The influence of DNA intercalator methylene blue on the impedance of both, single and double strands, was examined along with K-3[Fe(CN)(6)] and confirmed by cyclic voltammetry. The layer densities and the hybridisation have been further corroborated by chronoamperometric redox recycling of pal-a-aminophenol (p-AP) in ELISA like experiments. It can be concluded, that a performed impedance spectroscopy did not change the layer density. The impedance spectroscopy at ultramicroelectrodes combined with faradaic redox reactions enhances the impedimetric detection of DNA hybridisation on IDA platforms. (c) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:645 / 654
页数:10
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