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Use of the 16S-23S ribosomal genes spacer region for the molecular typing of sphingomonads

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The ability of sphingomonads in drinking water to cause community- and hospital-acquired opportunistic infections has raised the need to establish reproducible identification assays. In this study, a total of 129 isolates recovered from drinking water with yellow- to orange-pigmented colonies were distributed among 10 biotypes on the basis of colony morphology. Polymorphisms, based on the amplification and restriction digestion of the intergenic transcribed spacer (ITS) region within the 10 assigned biotypes and 18 ATCC reference strains, were used to investigate the ability of this approach to differentiate closely related sphingomonads. ITS size, which ranged between 400 and 1100 bp, did not vary enough among the different genera. However, 16 distinct banding patterns within the ATCC reference strains and 9 within the 10 biotypes were obtained through ITS restriction digestion, and the majority of the tested biotypes produced patterns similar to those generated by the ATCC strains. To our knowledge, this study is not only the first comprehensive record of the size of the ITS region in sphingomonads, it is also the first study that describes the use of ITS restriction digestion to subtype those isolates.

La capacité des sphingomonades de l’eau potable de causer des infections opportunistes acquises dans la communauté et dans les hôpitaux a mis en évidence le besoin d’établir des essais d’identification reproductibles. Dans cette étude, un total de 129 isolats recueillis de l’eau potable et comportant des colonies pigmentées de jaune à orange ont été séparés en 10 biotypes selon la morphologie des colonies. Le polymorphisme basé sur l’amplification et la digestion-restriction de la région de l’espaceur intergénique transcrit (ITS) des 10 biotypes assignés et de 18 souches de référence de l’ATCC a été utilisé pour examiner l’applicabilité de cette approche pour différencier des sphingomonades étroitement reliées. La taille de l’ITS, qui s’échelonnait de 400 à 1 100 pb, ne variait pas suffisamment entre les différents genres. Cependant, 16 patrons de digestion distincts ont été obtenus par digestion-restriction de l’ITS chez les souches de référence de l’ATCC et chez 9 des 10 biotypes, la majorité des biotypes présentant des patrons similaires à ceux générés avec les souches de l’ATCC. À notre connaissance, cette étude ne constitue pas seulement le premier rapport exhaustif des tailles de la région de l’ITS des sphingomonades, mais est aussi la première à décrire l’utilisation de la digestion-restriction de l’ITS pour typer ces isolats.

Document Type: Research Article

Publication date: 2008-08-01

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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