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Simultaneous Oligonucleotide Probe Hybridization and Immunostaining for in Situ Detection of Gordona Species in Activated Sludge
FEMS Microbiology Ecology
  • Daniel B. Oerther, Missouri University of Science and Technology
  • Francis L. de los Reyes, III
  • Mark Thomas Hernández
  • Lutgarde M. Raskin

Oligonucleotide probes targeting ribosomal RNA can be designed with high specificity to target microbial populations at different phylogenetic levels if the cellular abundance of ribosomal RNA is sufficiently high. In contrast, polyclonal antibody probes cannot be produced with the same specificity for various phylogenetic groups, but they have the potential to detect slow growing microorganisms, populations with low metabolic activities, or even non-viable cells. We combined a polyclonal antibody stain with a ribosomal RNA targeted oligonucleotide probe for the single cell detection of species of the genus Gordona. Gordona species typically require long generation times, often exhibit filamentous growth, and are commonly encountered in activated sludge foams. Our results suggest that the ribosomal RNA content of individual cells of Gordona in activated sludge is highly variable. Therefore, the combined use of an immunostain and an oligonucleotide probe targeting ribosomal RNA can determine the identity of single cells and provide an approximation of their activity. This approach should result in improved detection limits while maintaining high specificity.

Civil, Architectural and Environmental Engineering
Keywords and Phrases
  • polyclonal antibody,
  • ribosome RNA,
  • activated sludge,
  • methodology,
  • microbial community,
  • activated sludge,
  • controlled study,
  • fungus growth,
  • gordona,
  • Gordonia amarae,
  • immunohistochemistry,
  • nonhuman,
  • oligonucleotide probe,
  • priority journal,
  • RNA hybridization,
  • Fungi,
  • Gordonia (actinomycete),
  • Gordonia amarae,
  • Nocardia
Document Type
Article - Journal
Document Version
File Type
© 1999 Wiley-Blackwell, All rights reserved.
Publication Date
Citation Information
Daniel B. Oerther, Francis L. de los Reyes, Mark Thomas Hernández and Lutgarde M. Raskin. "Simultaneous Oligonucleotide Probe Hybridization and Immunostaining for in Situ Detection of Gordona Species in Activated Sludge" FEMS Microbiology Ecology Vol. 29 Iss. 2 (1999) p. 129 - 136 ISSN: 0168-6496
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