Simultaneous detection of dual single-base mutations by capillary electrophoresis using quantum dot-molecular beacon probe

Biosens Bioelectron. 2011 Jan 15;26(5):2317-22. doi: 10.1016/j.bios.2010.10.002. Epub 2010 Nov 5.

Abstract

Here a novel capillary electrophoresis (CE) for simultaneous detection of dual single-base mutations using quantum dot-molecular beacon (QD-MB) probe is described. Two QD-MB probes were designed using 585 and 650-nm emitting CdTe QDs which were covalently conjugated to MBs with different DNA oligonucleotide sequences by amide linkage and streptavidin-biotin binding, respectively. The hybridizations of QD-MB probes with different DNA targets were then monitored by CE, and results indicated that the two QD-MB probes specifically hybridized with their complementary DNA sequences, respectively. Target DNA identification was observed to have a high sensitivity of 16.2 pg in CE. Furthermore, the simultaneous detection of dual single-base mutations in a given DNA oligonucleotide was successfully achieved in CE using above two QD-MB probes. This novel CE-assisted QD-MB biosensor offers a promising approach for simultaneous detection of multiple single-base mutations, and exhibits potential capability in the single nucleotide polymorphism (SNP) analysis and high-sensitivity DNA detection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Pair Mismatch
  • Biosensing Techniques / instrumentation
  • DNA Mutational Analysis / instrumentation*
  • DNA Probes / genetics
  • Electrophoresis, Capillary / instrumentation*
  • Equipment Design
  • Equipment Failure Analysis
  • Microscopy, Fluorescence / instrumentation
  • Molecular Probe Techniques / instrumentation*
  • Polymorphism, Single Nucleotide / genetics*
  • Quantum Dots*
  • Sequence Analysis, DNA / instrumentation*

Substances

  • DNA Probes