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Titre :
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The domain-specific probe EUB338 is insufficient for the detection of all bacteria : Development and evaluation of a more comprehensive probe set
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Auteurs :
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H. Daims ;
A. Bruhl ;
R. Amann ;
AL Et
|
|
Type de document :
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article/chapitre/communication
|
|
Année de publication :
|
1999
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Format :
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p. 434-444
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Langues:
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= Anglais
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Catégories :
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Biologie
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Mots-clés:
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SONDE NUCLEIQUE
;
HYBRIDATION
;
ECOLOGIE BACTERIENNE
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Résumé :
|
In situ hybridization with rRNA-targeted oligonucleotide probes has become a widely applied tool for direct analysis of microbial population structures of complex natural and engineered systems. In such studies probe EUB338 (AMANN et al., 1990) is routinely used to quantify members of the domain Bacteria with a sufficiently high cellular ribosome content. Recent reevaluations of probe EUB338 coverage based on all publicly available 16S rRNA sequences, however, indicated that important bacterial phyla, most notably the Planctomycetales and Verrucomicrobia, are missed by this probe. We therefore designed and evaluated two supplementary versions (EUB338-II and EUB338-III) of probe EUB338 for in situ detection of most of those phyla not detected with probe EUB338. In situ dissociation curves with target and non-target organisms were recorded under increasing stringency to optimize hybridization conditions. For that purpose a digital image software routine was developed. In situ hybridization of a complex biofilm community with the three EUB338 probes demonstrated the presence of significant numbers of probe EUB338-II and EUB338-III target organisms.
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Source :
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Systematic and applied microbiology, vol.22
|
