Conditions of polymerase chain reaction amplification by magnetic enrichment and nanoscale detection sensitivity

J Nanosci Nanotechnol. 2012 May;12(5):3862-6. doi: 10.1166/jnn.2012.5868.

Abstract

In this research, we reported a method of polymerase chain reaction (PCR) amplification by means of magnetic enrichment. First, after denaturation, the target sequence was combined with biotin-modified specific primer through hybridization and enriched at the surface of gamma-Fe2O3 by biotin-avidin special interaction. Then single target sequence was gained through denaturation, and general PCR amplification was performed. The experiment conditions such as the hybridization temperature between target sequence and biotin-modified specific primer, and the dosage of magnetic nanoparticles gamma-Fe2O3 were optimized. Finally, the sensitivity of the method was checked. The lowest concentration of target sequence was detected as low as 5 x 10(-7) ng/mL. This simple method could provide a quick and early diagnosis of malignant infectious diseases such as SARS, avian flu and swine flu etc., that occur occasionally nowadays.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biotin / chemistry
  • DNA / chemistry
  • Ferric Compounds / chemistry*
  • Humans
  • Magnetics / methods*
  • Nanoparticles / chemistry*
  • Nanotechnology / methods
  • Nucleic Acid Hybridization / methods
  • Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Temperature

Substances

  • Ferric Compounds
  • Biotin
  • DNA