Enzyme-assisted target recycling (EATR) for nucleic acid detection

Chem Soc Rev. 2014 Sep 7;43(17):6405-38. doi: 10.1039/c4cs00083h.

Abstract

Fast, reliable and sensitive methods for nucleic acid detection are of growing practical interest with respect to molecular diagnostics of cancer, infectious and genetic diseases. Currently, PCR-based and other target amplification strategies are most extensively used in practice. At the same time, such assays have limitations that can be overcome by alternative approaches. There is a recent explosion in the design of methods that amplify the signal produced by a nucleic acid target, without changing its copy number. This review aims at systematization and critical analysis of the enzyme-assisted target recycling (EATR) signal amplification technique. The approach uses nucleases to recognize and cleave the probe-target complex. Cleavage reactions produce a detectable signal. The advantages of such techniques are potentially low sensitivity to contamination and lack of the requirement of a thermal cycler. Nucleases used for EATR include sequence-dependent restriction or nicking endonucleases or sequence independent exonuclease III, lambda exonuclease, RNase H, RNase HII, AP endonuclease, duplex-specific nuclease, DNase I, or T7 exonuclease. EATR-based assays are potentially useful for point-of-care diagnostics, single nucleotide polymorphisms genotyping and microRNA analysis. Specificity, limit of detection and the potential impact of EATR strategies on molecular diagnostics are discussed.

Publication types

  • Review

MeSH terms

  • Base Sequence
  • DNA / analysis*
  • DNA / chemistry
  • DNA / genetics
  • DNA / metabolism
  • DNA Restriction Enzymes / metabolism*
  • Humans
  • Nucleic Acid Amplification Techniques / methods*
  • Nucleic Acid Hybridization
  • Oligonucleotide Probes / chemistry
  • Oligonucleotide Probes / genetics
  • RNA / analysis*
  • RNA / chemistry
  • RNA / genetics
  • RNA / metabolism

Substances

  • Oligonucleotide Probes
  • RNA
  • DNA
  • DNA Restriction Enzymes