Highly efficient isothermal DNA amplification system using three elements of 5'-DNA-RNA-3' chimeric primers, RNaseH and strand-displacing DNA polymerase

J Biochem. 2007 Aug;142(2):273-81. doi: 10.1093/jb/mvm138. Epub 2007 Aug 24.


We developed an efficient method of isothermally amplifying DNA termed ICAN, Isothermal and Chimeric primer-initiated Amplification of Nucleic acids. This method allows the amplification of target DNA under isothermal conditions at around 55 degrees C using only a pair of 5'-DNA-RNA-3' chimeric primers, a thermostable RNaseH and a DNA polymerase with strong strand-displacing activity. ICAN is capable of amplifying DNA at least several times greater than the amount produced with PCR by increasing primer concentration. This method would be applicable for on-site DNA detection including gene diagnosis, and would also be suitable for 'real time' detection when combined with a cycling probe.

Publication types

  • Evaluation Study

MeSH terms

  • DNA / metabolism*
  • DNA Primers / chemistry*
  • DNA-Directed DNA Polymerase / metabolism*
  • Nucleic Acid Amplification Techniques / methods*
  • RNA / chemistry*
  • Ribonuclease H / chemistry
  • Ribonuclease H / metabolism*
  • Temperature*


  • DNA Primers
  • RNA primers
  • RNA
  • DNA
  • DNA-Directed DNA Polymerase
  • Ribonuclease H