Catalyzed reporter deposition-fluorescent in situ hybridization (CARD-FISH) detection of Dehalococcoides

Journal of Microbiological Methods
J A DijkD Springael

Abstract

Members of the genus Dehalococcoides are well-known for their capacity to reductively dechlorinate chlorinated organic pollutants. The availability of quantitative and sensitive detection methods is of major interest for research on the ecology of those environmentally important micro-organisms. In this paper we describe the development of a Catalyzed Reporter Deposition-Fluorescent In Situ Hybridization (CARD-FISH) for detection of Dehalococcoides cells in enrichment cultures using two oligonucleotide sequences which target two different lineages of Dehalococcoides as probes. Both sequences were previously applied in conventional FISH as probes. Conjugation of the probe to horseradish peroxidase (HRP) did not change the specificity of the probes and bright fluorescent signals were obtained. Despite the use of higher concentrations of probe and the application of longer exposure times in the conventional FISH procedure, CARD-FISH resulted in more intense signals. The CARD-FISH method was applied to a vinyl chloride (VC)-reductively-dechlorinating enrichment culture. Only the probe targeting the CBDB1 lineage of Dehalococcoides reacted with the sample which was in agreement with previous nucleic acid based analysis. The culture ...Continue Reading

References

Dec 20, 1989·Journal of Immunological Methods·M N BobrowG J Litt
Jun 1, 2002·Applied and Environmental Microbiology·Annelie PernthalerRudolf Amann
Sep 4, 2004·Applied and Environmental Microbiology·Tina HölscherLorenz Adrian
Sep 4, 2004·Applied and Environmental Microbiology·Melanie DuhamelElizabeth A Edwards
Sep 15, 2005·Environmental Microbiology·Stefano FaziAlberto Puddu
Oct 12, 2005·Journal of Microbiological Methods·Niina A TujulaGregory R Crocetti
Mar 7, 2006·Applied and Environmental Microbiology·Youlboong SungFrank E Löffler
Apr 7, 2006·Applied and Environmental Microbiology·Kirsti M RitalahtiFrank E Löffler

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Citations

Oct 19, 2010·Journal of Contaminant Hydrology·P J HaestE Smolders
Oct 19, 2010·Analytica Chimica Acta·Jason WojciechowskiChris Rowe Taitt
May 29, 2010·FEMS Microbiology Ecology·Danuta CichockaIvonne Nijenhuis

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