Electrochemical detection of the toxic dinoflagellate Alexandrium ostenfeldii with a DNA-biosensor

Biosens Bioelectron. 2005 Jan 15;20(7):1349-57. doi: 10.1016/j.bios.2004.05.011.

Abstract

The steady rise of observations of harmful or toxic algal blooms throughout the world in the past decades constitute a menace for coastal ecosystems and human interests. As a consequence, a number of programs have been launched to monitor the occurrence of harmful and toxic algae. However, the identification is currently done by microscopic examination, which requires a broad taxonomic knowledge, expensive equipment and is very time consuming. In order to facilitate the identification of toxic algae, an inexpensive and easy-to-handle DNA-biosensor has been adapted for the electrochemical detection of the toxic dinoflagellate Alexandrium ostenfeldii. The detection of the toxic algae is based on a sandwich hybridisation, which is carried out on a disposable sensor chip. A set of two probes for the species-specific identification of A. ostenfeldii was developed. The specificity of the probes could be shown in dot-blot hybridisations and with the DNA-biosensor. The sensitivity of the DNA-biosensor was optimised with respect to hybridisation temperature and NaCl-concentration and a significant increase of the sensitivity of the DNA-biosensor could be obtained by a fragmentation of the rRNA prior to the hybridisation and by adding a helper oligonucleotide, which binds in close proximity to the probes to the hybridisation.

MeSH terms

  • Animals
  • Biosensing Techniques*
  • DNA*
  • Dinoflagellida / genetics
  • Dinoflagellida / isolation & purification*
  • Humans
  • Molecular Probes
  • RNA, Ribosomal

Substances

  • Molecular Probes
  • RNA, Ribosomal
  • DNA