This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Rudi, K.
Right arrow Articles by Jakobsen, K. S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Rudi, K.
Right arrow Articles by Jakobsen, K. S.
Agricola
Right arrow Articles by Rudi, K.
Right arrow Articles by Jakobsen, K. S.

 Previous Article  |  Next Article 

Appl Environ Microbiol, January 1998, p. 34-37, Vol. 64, No. 1
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Detection of Toxin-Producing Cyanobacteria by Use of Paramagnetic Beads for Cell Concentration and DNA Purification

Knut Rudi,1,* Frank Larsen,2 and Kjetill S. Jakobsen1

Division of General Genetics, Department of Biology, University of Oslo, 0315 Oslo,1 and Dynal A.S., 0212 Oslo,2 Norway

Received 13 August 1997/Accepted 17 October 1997

Early detection of water blooms caused by potential toxin-producing cyanobacteria is important in environmental monitoring. We present a new nucleic acid-based method for detection of cyanobacteria in water that utilizes the same paramagnetic solid phase (beads) for both bacterial cell concentration and subsequent DNA purification. In the cell concentration step, the beads were attracted to a magnet after cell adsorption (in an alcohol- and salt-containing solution), and the supernatant was removed. For DNA purification, a buffer containing guanidine thiocyanate and Sarkosyl lysed the concentrated cells. The addition of alcohol precipitated the released DNA onto the same solid phase as was used for the cell concentration. Finally, to remove PCR inhibitors, the DNA was washed twice in alcohol while bound to the beads. All of the bead-DNA complex was used in the subsequent PCR amplification. The detection limit, as measured by 16S rDNA PCR amplification, was 50 cells in a 0.5-ml water sample, which is considerably lower than the limit (500 cells/ml) of toxic cyanobacteria tolerated in drinking water (New South Wales Blue-Green Algae Task Force, 1992). Testing of water from natural habitats showed a detection limit in the same range as that for the defined samples. The detection limits and the simplicity of the method (paramagnetic beads can be handled in automated systems) suggest that our method is suitable for routine environmental monitoring.

* Corresponding author. Mailing address: Division of General Genetics, Department of Biology, University of Oslo, P.O. Box 1031 Blindern, 0315 Oslo, Norway. Phone: (47.)22.85.45.73. Fax: (47.)22.85.46.05. E-mail: knut.rudi{at}bio.uio.no.




This article has been cited by other articles:

  • Hotto, A. M., Satchwell, M. F., Boyer, G. L. (2007). Molecular Characterization of Potential Microcystin-Producing Cyanobacteria in Lake Ontario Embayments and Nearshore Waters. Appl. Environ. Microbiol. 73: 4570-4578 [Abstract] [Full Text]  
  • Rudi, K., Hoidal, H. K., Katla, T., Johansen, B. K., Nordal, J., Jakobsen, K. S. (2004). Direct Real-Time PCR Quantification of Campylobacter jejuni in Chicken Fecal and Cecal Samples by Integrated Cell Concentration and DNA Purification. Appl. Environ. Microbiol. 70: 790-797 [Abstract] [Full Text]  
  • Nogva, H. K., Rudi, K., Naterstad, K., Holck, A., Lillehaug, D. (2000). Application of 5'-Nuclease PCR for Quantitative Detection of Listeria monocytogenes in Pure Cultures, Water, Skim Milk, and Unpasteurized Whole Milk. Appl. Environ. Microbiol. 66: 4266-4271 [Abstract] [Full Text]  
  • Rudi, K., Skulberg, O. M., Skulberg, R., Jakobsen, K. S. (2000). Application of Sequence-Specific Labeled 16S rRNA Gene Oligonucleotide Probes for Genetic Profiling of Cyanobacterial Abundance and Diversity by Array Hybridization. Appl. Environ. Microbiol. 66: 4004-4011 [Abstract] [Full Text]  
  • Fergusson, K. M., Saint, C. P. (2000). Molecular Phylogeny of Anabaena circinalis and Its Identification in Environmental Samples by PCR. Appl. Environ. Microbiol. 66: 4145-4148 [Abstract] [Full Text]  
  • Rudi, K., Skulberg, O. M., Larsen, F., Jakobsen, K. S. (1998). Quantification of Toxic Cyanobacteria in Water by Use of Competitive PCR Followed by Sequence-Specific Labeling of Oligonucleotide Probes. Appl. Environ. Microbiol. 64: 2639-2643 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»