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


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kmetfies [ at ] awi-bremerhaven.de

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 hybridisation 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.



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Published
Eprint ID
11280
DOI 10.1016/j.bios.2004.05.011

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Metfies, K. , Huljic, S. , Lange, M. and Medlin, L. (2005): Electrochemical detection of the toxic dinoflagellate Alexandrium ostenfeldii with a DNA-biosensor , Biosensors & bioelectronics, 20 , pp. 1349-1357 . doi: 10.1016/j.bios.2004.05.011


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