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Applied and Environmental Microbiology, January 2006, p. 918-922, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.918-922.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Catalyzed Reporter Deposition-Fluorescence In Situ Hybridization Allows for Enrichment-Independent Detection of Microcolony-Forming Soil Bacteria

Belinda C. Ferrari,1* Niina Tujula,2 Kate Stoner,1 and Staffan Kjelleberg2

Department of Chemistry and Biomolecular Sciences, Division of Environmental and Life Sciences, Macquarie University, Sydney, Australia,1 Centre for Marine Biofouling and Bio-Innovation, School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, New South Wales, Australia2

Received 1 July 2005/ Accepted 1 November 2005

Advances in the growth of hitherto unculturable soil bacteria have emphasized the requirement for rapid bacterial identification methods. Due to the slow-growing strategy of microcolony-forming soil bacteria, successful fluorescence in situ hybridization (FISH) requires an rRNA enrichment step for visualization. In this study, catalyzed reporter deposition (CARD)-FISH was employed as an alternative method to rRNA enhancement and was found to be superior to conventional FISH for the detection of microcolonies that are cultivated by using the soil substrate membrane system. CARD-FISH enabled real-time identification of oligophilic microcolony-forming soil bacteria without the requirement for enrichment on complex media and the associated shifts in community composition.

* Corresponding author. Mailing address: Department of Chemistry and Biomolecular Sciences, Division of Environmental and Life Sciences, Macquarie University, Sydney, NSW 2109, Australia. Phone: 61 2 9850 9252. Fax: 612 9850 8253. E-mail: bferrari{at}rna.bio.mq.edu.au

Applied and Environmental Microbiology, January 2006, p. 918-922, Vol. 72, No. 1
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.1.918-922.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Ferrari, B. C., Gillings, M. R. (2009). Cultivation of Fastidious Bacteria by Viability Staining and Micromanipulation in a Soil Substrate Membrane System. Appl. Environ. Microbiol. 75: 3352-3354 [Abstract] [Full Text]  


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