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JPR Advance Access published online on January 29, 2008
Journal of Plankton Research, doi:10.1093/plankt/fbn009
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© The Author 2008. Published by Oxford University Press. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Molecular probe sets for the detection of toxic algae for use in sandwich hybridisation formats

Sonja Diercks*, Katja Metfies and Linda K. Medlin

Alfred Wegener Institute for Polar and Marine Research, Am Handelshafen 12, 27570 Bremerhaven, Germany

* Corresponding author: Phone: +49 471 4831 1384 Fax: +49 471 4831 1425 E-mail: Sonja.Diercks{at}awi.de Alfred Wegener Institute for Polar and Marine Research, Am Handelshafen 12, 27570 Bremerhaven, Germany

Received on February 26, 2007; revised on January 14, 2008; accepted on January 16, 2008


  Abstract

Molecular probes can be used for early and rapid detection of toxic algal species. The sandwich hybridisation requires two probes for each species, a capture probe and a nearly adjacent signal probe. Probe sets for the species-specific identification of the toxic algal species Gymnodinium catenatum, Protoceratium reticulatum, Lingulodinium polyedrum, Prymnesium parvum, Chrysochromulina polylepis, Pseudo-nitzschia multiseries, P. australis, P. seriata, and P. pungens were designed. A genus probe set for Pseudo-nitzschia species was adapted, and all probe sets were tested for specificity. The target molecules for the probe sets are the large and the small subunit ribosomal RNAs. The specificity of the different probe sets was tested using a sandwich hybridisation assay (SHA) in a microtiter plate with ribosomal RNA isolated from laboratory strains of the target species and closely related species. The assay showed that eight probe sets were highly specific. For Gymnodinium catenatum and Prymnesium parvum, the probe sets detected one non-target species outside the target species. These ten probe sets are valuable tools for identifying and monitoring different toxic algae. The microtiter plate assay is a cheap and effective means for testing probe specificity.

Key Words: DNA probes • Microtiter plate assay • Sandwich hybridisation • Toxic algae

Communicating Editor: Dr Roger Harris


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