A cyclic enzymatic amplification method for sensitive and selective detection of nucleic acids

Analyst. 2010 Aug;135(8):2069-73. doi: 10.1039/c0an00215a. Epub 2010 Jun 18.

Abstract

Based on Exonuclease III (Exo III) and displacing probes, we have developed a Cyclic Enzymatic Amplification Method (CEAM) for sensitive and selective detection of nucleic acids. In this design, the displacing probe is non-fluorescent on its own and cannot be digested by Exo III until displacement hybridization by a target sequence, leading to release of free non-quenched fluorophore. Because a single target sequence can lead to the release and digestion of numerous fluorophore strands from the displacing probe, a remarkable signal amplification is achieved. With this method, DNA can be detected in the picomolar range with a high selectivity and within less than 20 min.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Exodeoxyribonucleases / chemistry
  • Exodeoxyribonucleases / metabolism*
  • Nucleic Acid Amplification Techniques / methods*
  • Nucleic Acids / genetics*
  • Time Factors

Substances

  • Nucleic Acids
  • Exodeoxyribonucleases
  • exodeoxyribonuclease III