Studies on the fluorescence fiber-optic DNA biosensor using p-hydroxyphenylimidazo[f]1,10-phenanthroline ferrum(III) as indicator

J Fluoresc. 2008 Jan;18(1):227-35. doi: 10.1007/s10895-007-0266-1. Epub 2007 Oct 16.

Abstract

A complex Fe(phen)(2).PHPIP.3ClO(4).2H(2)O, where phen=1,10-phenanthroline and PHPIP=p-hydroxyphenylimidazo[f]1,10-phenanthroline, was synthesized and acted as a good fluorescence indicator based on its interaction with double-duplex DNA. Then a fiber-optic DNA biosensor of fluorimetric detection was developed based on the recognition of target DNA in DNA hybridization assays. A probe ssDNA was covalently immobilized onto the surface of quartz optical fibers and then the probe ssDNA hybridized with complementary ssDNA introduced into the local environment of the sensor. The hybridization with complementary strands was monitored in real time by fluorimetric detection. Several factors affecting the probe immobilization, target DNA hybridization, and indicator binding reactions were optimized to maximize the sensitivity and shorten the assay time. Using this method, a sequence of the 16-mer oligonucleotides could be quantified over the range from 4.98 x 10(-7) to 4.88 x 10(-6) M and a detection limit of 1.08 x 10(-7) M. And the designed optic-fiber biosensor could be conveniently regenerated by thermal denature. The utility of the novel hybridization indicator could provide a simple, rapid, low toxicity and reusable detection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques*
  • DNA, Single-Stranded / chemistry*
  • DNA, Single-Stranded / metabolism*
  • Ferric Compounds / chemical synthesis
  • Ferric Compounds / chemistry
  • Ferric Compounds / metabolism*
  • Fiber Optic Technology*
  • Fluorescence
  • Nucleic Acid Hybridization
  • Optical Fibers
  • Phenanthrolines / chemical synthesis
  • Phenanthrolines / chemistry
  • Phenanthrolines / metabolism*
  • Spectrometry, Fluorescence

Substances

  • DNA, Single-Stranded
  • Ferric Compounds
  • Phenanthrolines
  • p-hydroxyphenylimidazo(f)1,10-phenanthroline ferrum(III)