Detection of single-nucleotide polymorphism on uidA gene of Escherichia coli by a multiplexed electrochemical DNA biosensor with oligonucleotide-incorporated nonfouling surface

Sensors (Basel). 2011;11(8):8018-27. doi: 10.3390/s110808018. Epub 2011 Aug 15.

Abstract

We report here a practical application of a multiplexed electrochemical DNA sensor for highly specific single-nucleotide polymorphism (SNP) detection. In this work, a 16-electrode array was applied with an oligonucleotide-incorporated nonfouling surfaces (ONS) on each electrode for the resistance of unspecific absorption. The fully matched target DNA templated the ligation between the capture probe assembled on gold electrodes and the tandem signal probe with a biotin moiety, which could be transduced to peroxidase-based catalyzed amperometric signals. A mutant site (T93G) in uidA gene of E. coli was analyzed in PCR amplicons. 10% percentage of single mismatched mutant gene was detected, which clearly proved the selectivity of the multiplexed electrochemical DNA biosensor when practically applied.

Keywords: Escherichia coli; electrochemical biosensor; nonfouling electrode surface; single-nucleotide polymorphism (SNP).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Biosensing Techniques / methods*
  • Biotin
  • DNA / chemistry*
  • DNA / genetics
  • Electrochemistry / methods*
  • Electrodes
  • Escherichia coli Proteins / genetics*
  • Glucuronidase / genetics*
  • Gold / chemistry
  • Molecular Sequence Data
  • Mutation
  • Oligonucleotides / chemistry*
  • Peroxidase / chemistry
  • Polymerase Chain Reaction / methods
  • Polymorphism, Single Nucleotide*

Substances

  • Escherichia coli Proteins
  • Oligonucleotides
  • Biotin
  • Gold
  • DNA
  • Peroxidase
  • Glucuronidase