A detection method for Legionella spp in (cooling) water: fluorescent in situ hybridisation (FISH) on whole bacteria

2003 ◽  
Vol 47 (3) ◽  
pp. 143-146 ◽  
Author(s):  
P. Declerck ◽  
L. Verelst ◽  
L. Duvivier ◽  
A. Van Damme ◽  
F. Ollevier
Keyword(s):  
16S Rrna ◽  
In Situ Hybridisation ◽  
Cooling Water ◽  
Culture Methods ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Fluorescent In Situ Hybridisation ◽  
Background Staining ◽  
Legionella Species

Although traditional culture methods are appropriate for detection of Legionella species, such culture takes several days. Rapid detection (<24 h) of individual Legionella is possible using fluorescent in situ hybridisation (FISH) on whole bacteria. Water samples were filtered and the concentrated bacteria were immediately detected (without culture) with a fluorescence microscope following appropriate labelling. The detection level was very high and quantification was possible. For the detection of all Legionella spp. the probe LEG705 was used, complementary to a 16S rRNA sequence conserved in all Legionella spp. For specific detection of L. pneumophila the probe LEGPNE1 was used. This probe is designed against a variable domain of the 16S rRNA sequence from L. pneumophila. CY3 and FLUOS labels were tested and CY3 showed clearly detectable bacteria with minimum background staining. This FISH technique is very sensitive, fast, reliable and individual bacteria are easily detected.

scholarly journals In Situ Detection of an Uncultured Predominant Bacillus in Dutch Grassland Soils

1998 ◽  
Vol 64 (11) ◽  
pp. 4588-4590 ◽  
Author(s):  
Andreas Felske ◽  
Antoon D. L. Akkermans ◽  
Willem M. De Vos
Keyword(s):  
In Situ Hybridization ◽  
16S Rrna ◽  
Cell Type ◽  
Grassland Soils ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Whole Cell ◽  
Shaped Cell ◽  
In Situ Detection

ABSTRACT Uncultured predominant Bacillus ribotype DA001 in Dutch Drentse A grassland soils, as revealed by its 16S rRNA sequence, was detected in soil by fluorescent whole-cell in situ hybridization. A prominent rod-shaped cell type was identified in bacterial suspensions prepared from soil by a multiple 16S rRNA probing approach.

scholarly journals Phylogeny and Identification In Situ ofNevskia ramosa

1998 ◽  
Vol 64 (5) ◽  
pp. 1895-1901 ◽  
Author(s):  
Frank Oliver Gl�ckner ◽  
Hans-Dietrich Babenzien ◽  
Rudolf Amann
Keyword(s):  
16S Rrna ◽  
Cloning And Sequencing ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Bacterial Populations ◽  
Freshwater Habitats ◽  
Related Group ◽  
Rrna Sequences ◽  
16S Rrna Sequences

ABSTRACT An enrichment of the neuston bacterium Nevskia ramosawas investigated by the cultivation-independent rRNA approach. N. ramosa was first described by Famintzin in 1892 as a rod-shaped, slightly bent bacterium forming typical flat rosettes on the surface of shallow freshwater habitats by unilateral slime formation. PCR in combination with cloning and sequencing was used for retrieving 21 partial and 5 nearly full-length 16S rRNA sequences forming three tight clusters. In situ hybridization with rRNA-targeted oligonucleotide probes allowed us to assign the three sequence clusters to three distinct bacterial populations abundant in the enrichment. The two probes that unambiguously identified the N. ramosamorphotype were derived from a 16S rRNA sequence that had similarities of 87.9 to 88.9% to the rRNA sequences of the most closely related group in the database, Xanthomonas sp. and relatives.N. ramosa currently is the only representative of an independent, deep branch of the gamma subclass of the classProteobacteria. The two other populations abundant in the enrichment were affiliated with the alpha subclass of the classProteobacteria. They were most closely related toBlastobacter sp. (97.2% similarity) and Mycoplana bullata (97.6% similarity) and might represent new species in the respective genera.

scholarly journals 16S rRNA Sequence-based Rapid and Sensitive Detection of Aquatic Bacteria by On-chip Hybridization Following Multiplex PCR

2008 ◽  
Vol 54 (1) ◽  
pp. 123-128 ◽  
Author(s):  
Tomoaki Ichijo ◽  
Nobuyasu Yamaguchi ◽  
Katsuji Tani ◽  
Masao Nasu
Keyword(s):  
16S Rrna ◽  
Multiplex Pcr ◽  
Sensitive Detection ◽  
Rrna Sequence ◽  
16S Rrna Sequence ◽  
Aquatic Bacteria ◽  
On Chip
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