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Detection of short oligonucleotide sequences of hepatitis B virus using electrochemical DNA hybridisation biosensor

Sophia Karastogianni and Stella Girousi

Department of Analytical Chemistry, Aristotle University of Thessaloniki, Panepistimioupoli Thessaloniki 54124, Greece

 

E-mail: girousi@chem.auth.gr

Abstract: A novel, sensitive and selective electrochemical hybridisation biosensor was developed for the detection of the hepatitis B virus (HBV) using a manganese(II) complex as electrochemical indicator and a DNA probe-modified carbon paste electrode as the biosensor (DNA/CPE). The results showed that this complex could be accumulated electrochemically the immobilised dsDNA layer rather than in the single-stranded DNA (ssDNA) layer. On the basis of this, the manganese complex was used as an electrochemical hybridisation indicator for the detection of oligonucleotides related to HBV. The hybridisation event was evaluated on the basis of the difference between the reduction signals of the manganese(II) complex with the probe DNA prior to and post hybridisation with a target sequence using a differential pulse mode. Several factors affecting the immobilisation and hybridisation of oligonucleotides as well as the indicator’s accumulation were investigated. Experiments with a noncomplementary and mismatch sequences demonstrated the good selectivity of the biosensor. Using this approach, the HBV target oligonucleotide’s sequence could be quantified over arange from 0.22 ng L−1 to 5.40 ng L−1, with a linear correlation coefficient of 0.9994 and the limit of detection of 0.07 ng L−1.

Keywords: Mn(II) complex – differential pulse voltammetry – DNA-hybridisation biosensor – oligonucleotides

Full paper is available at www.springerlink.com.

DOI: 10.2478/s11696-014-0599-6

 

Chemical Papers 69 (1) 202–210 (2015)

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