Kukol, A and Li, P and Estrela, P. M. and Ko-Ferrigno, P and Migliorato, P (2008) Label-free electrical detection of DNA hybridization for the example of influenza virus gene sequences. Analytical Biochemistry, 374 (1). pp. 143-153. ISSN 0003-2697
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Official URL: http://publications.eng.cam.ac.uk/14931/
Microarrays based on DNA–DNA hybridization are potentially useful for detecting and subtyping viruses but require fluorescence labeling and imaging equipment. We investigated a label-free electrical detection system using electrochemical impedance spectroscopy that is able to detect hybridization of DNA target sequences derived from avian H5N1 influenza virus to gold surface-attached single-stranded DNA oligonucleotide probes. A 23-nt probe is able to detect a 120-nt base fragment of the influenza A hemagglutinin gene sequence. We describe a novel method of data analysis that is compatible with automatic measurement without operator input, contrary to curve fitting used in conventional electrochemical impedance spectroscopy (EIS) data analysis. A systematic investigation of the detection signal for various spacer molecules between the oligonucleotide probe and the gold surface revealed that the signal/background ratio improves as the length of the spacer increases, with a 12- to 18-atom spacer element being optimal. The optimal spacer molecule allows a detection limit between 30 and 100 fmol DNA with a macroscopic gold disc electrode of 1 mm radius. The dependence of the detection signal on the concentration of a 23-nt target follows a binding curve with an approximate 1:1 stoichiometry and a dissociation constant of KD = 13 ± 4 nM at 295 K.
|Uncontrolled Keywords:||DNA; Biosensor; Avian influenza; Label-free detection; Electrochemical impedance spectroscopy; Sulfhydryl chemistry; Spacer molecule; Oligonucleotide|
|Subjects:||Analytical Science > Microscopy and probe methods|
Biomedical Science > Nanoscale biological processes
|Deposited By:||Lesley Tobin|
|Deposited On:||02 Dec 2008 14:07|
|Last Modified:||25 Feb 2009 11:53|
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