Specific Oligonucleotide Probes for in situ Detection of a Major Group of Gram-positive Bacteria with low DNA G+C Content

doi:10.1016/S0723-2020(99)80065-4

Systematic and Applied Microbiology

Volume 22, Issue 2, May 1999, Pages 186-196

https://doi.org/10.1016/S0723-2020(99)80065-4 Get rights and content

Summary

Almost one thousand 16S rRNA sequences of Gram-positive bacteria with a low DNA G+C content from public databases were analyzed using the ARB software package. A signature region was identified between positions 354 and 371 (E. coli numbering) for the Bacillus sub-branch of the Gram-positive bacteria with a low DNA G+C content, the former orders Bacillales and Lactobacillales. Three oligonucleotide probes, namely LGC354A, LGC354B, and LGC354C, were designed to target this diagnostic site. Their fluorescent derivatives were suitable for whole cell detection by fluorescence in situ hybridization (FISH). Hybridization conditions were adjusted for differentiation of target and related non-target reference species. When applying FISH to whole bacterial cells in a sample of activated sludge from a communal wastewater treatment plant, members of the Bacillus sub-branch were detected at levels from 0.01% of cells in samples fixed with paraformaldehyde to over 8 percent in the same samples fixed with ethanol and treated with lysozyme. The problems of quantitative in situ analysis of Gram-positive bacteria with a low DNA G+C content in biofilm flocs are discussed and recommendations made. Members of the Bacillus sub-branch were detected in different abundances in activated sludge samples from different wastewater plants.

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Specific Oligonucleotide Probes for in situ Detection of a Major Group of Gram-positive Bacteria with low DNA G+C Content

Summary

System. Appl. Microbiol.

J. Molec. Biol.

System. Appl. Microbiol

System. Appl. Microbiol.

System. Appl. Microbiol.

System. Appl. Microbiol.

Int. J. Food Microbiol.

Community analysis of the bacterial assemblages in the winter cover and pelagic layers of a high mountain lake by in situ hybridization

Appl. Environ. Microbiol.

Fluorescentoligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology

J. Bacteriol.

Combination of 16S rRNA-targed oligonucleotide probes with flow cytometry for analyzing mixed microbial populations

Appl. Environ. Microbiol.

Phylogenetic identification and in situ detection of individual microbial cells without cultivation

Microbial Review

Phylogenetic stains: ribosomal RNA-based probes for the identification of single microbial cells

Science

Evolution according to large ribosomal subunit RNA J

Mol. Evol.

Evolutionary trees from DNA sequences: A maximum likelihood approach

J. Mol. Evol.

The phylogeny of prokaryotes

Science