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PMID: 2200342 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S.

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

Applied and environmental microbiology ·Vol. 56 ·No. 6 ·1990-06-00 ·Pages 1919-25

Amann RI, Binder BJ, Olson RJ, Chisholm SW, Devereux R, Stahl DA

Abstract

Fluorescent oligonucleotide hybridization probes were used to label bacterial cells for analysis by flow cytometry. The probes, complementary to short sequence elements within the 16S rRNA common to phylogenetically coherent assemblages of microorganisms, were labeled with tetramethylrhodamine and hybridized to suspensions of fixed cells. Flow cytometry was used to resolve individual target and nontarget bacteria (1 to 5 microns) via probe-conferred fluorescence. Target cells were quantified in an excess of nontarget cells. The intensity of fluorescence was increased additively by the combined use of two or three fluorescent probes complementary to different regions of the same 16S rRNA.

MeSH Terms
Base Sequence Escherichia coli/genetics Flow Cytometry Fluorescent Dyes Molecular Sequence Data Oligonucleotide Probes RNA, Ribosomal/genetics RNA, Ribosomal, 16S/genetics Ribosomes/analysis
Chemicals
Fluorescent Dyes Oligonucleotide Probes RNA, Ribosomal RNA, Ribosomal, 16S
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Amann R I
Department of Veterinary Pathobiology, University of Illinois, Urbana 61801.
Binder B J
Olson R J
Chisholm S W
Devereux R
Stahl D A
References (18)
18 references, click to expand
  1. Phylogenetic group-specific oligodeoxynucleotide probes for identification of single microbial cells.
    J Bacteriol. 1988 Feb;170(2):720-6 PMID: 2448289
  2. Natural relationships among sulfate-reducing eubacteria.
    J Bacteriol. 1989 Dec;171(12):6689-95 PMID: 2480344
  3. Complete nucleotide sequence of a 16S ribosomal RNA gene from Escherichia coli.
    Proc Natl Acad Sci U S A. 1978 Oct;75(10):4801-5 PMID: 368799
  4. Natural relationship between bacteroides and flavobacteria.
    J Bacteriol. 1985 Oct;164(1):230-6 PMID: 2413007
  5. A phylogenetic definition of the major eubacterial taxa.
    Syst Appl Microbiol. 1985;6:143-51 PMID: 11542017
  6. Use of phylogenetically based hybridization probes for studies of ruminal microbial ecology.
    Appl Environ Microbiol. 1988 May;54(5):1079-84 PMID: 3389805
  7. Phylogenetic stains: ribosomal RNA-based probes for the identification of single cells.
    Science. 1989 Mar 10;243(4896):1360-3 PMID: 2466341
  8. Marine ammonia- and nitrite-oxidizing bacteria: serological diversity determined by immunofluorescence in culture and in the environment.
    Appl Environ Microbiol. 1985 Aug;50(2):194-201 PMID: 16346845
  9. Use of nuclepore filters for counting bacteria by fluorescence microscopy.
    Appl Environ Microbiol. 1977 May;33(5):1225-8 PMID: 327932
  10. Bacterial evolution.
    Microbiol Rev. 1987 Jun;51(2):221-71 PMID: 2439888
  11. Conversion of logarithmic channel numbers into relative linear fluorescence intensity.
    Cytometry. 1988 Nov;9(6):533-8 PMID: 3264784
  12. Fluorescent-oligonucleotide probing of whole cells for determinative, phylogenetic, and environmental studies in microbiology.
    J Bacteriol. 1990 Feb;172(2):762-70 PMID: 1688842
  13. Identification and enumeration of marine chroococcoid cyanobacteria by immunofluorescence.
    Appl Environ Microbiol. 1983 Sep;46(3):553-9 PMID: 16346382
  14. Nucleotide sequence of the Bacillus subtilis ribosomal RNA operon, rrnB.
    Gene. 1985;37(1-3):261-6 PMID: 2414156
  15. Characterizing aquatic bacteria according to population, cell size, and apparent DNA content by flow cytometry.
    Cytometry. 1989 Jan;10(1):70-6 PMID: 2465113
  16. Flow cytometric detection of ribosomal RNA in suspended cells by fluorescent in situ hybridization.
    Cytometry. 1988 Nov;9(6):517-24 PMID: 3208617
  17. Shifts in methanogenic subpopulations measured with antibody probes in a fixed-bed loop anaerobic bioreactor treating sulfite evaporator condensate.
    Appl Environ Microbiol. 1989 Aug;55(8):1996-2001 PMID: 16347990
  18. Microbial ecology and evolution: a ribosomal RNA approach.
    Annu Rev Microbiol. 1986;40:337-65 PMID: 2430518
Article Info
Journal
Applied and environmental microbiology
Abbr.
Appl Environ Microbiol
ISSN
0099-2240
DOI
10.1128/aem.56.6.1919-1925.1990
Published
1990-06-00
Pages
1919-25
Language
English
Region
United States
NLM ID
7605801
PMCID
PMC184531
Subset
IM
External Links
PubMed source DOI: 10.1128/aem.56.6.1919-1925.1990 PMC Full Text View journal details
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