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Comparative Study
. 2001 Jan;39(1):381-4.
doi: 10.1128/JCM.39.1.381-384.2001.

Ralstonia paucula (Formerly CDC group IV c-2): unsuccessful strain differentiation with PCR-based methods, study of the 16S-23S spacer of the rRNA operon, and comparison with other Ralstonia species (R. eutropha, R. pickettii, R. gilardii, and R. solanacearum)

D Moissenet  1 , P Bidet, A Garbarg-Chenon, G Arlet, H Vu-Thien
Affiliations
Comparative Study

Ralstonia paucula (Formerly CDC group IV c-2): unsuccessful strain differentiation with PCR-based methods, study of the 16S-23S spacer of the rRNA operon, and comparison with other Ralstonia species (R. eutropha, R. pickettii, R. gilardii, and R. solanacearum)

D Moissenet et al. J Clin Microbiol. 2001 Jan.

Abstract

Ralstonia paucula (formerly CDC group IV c-2) can cause serious human infections. Confronted in 1995 with five cases of nosocomial bacteremia, we found that pulsed-field gel electrophoresis could not distinguish between the isolates and that randomly amplified polymorphic DNA analysis was poorly discriminatory. In this study, we used PCR-ribotyping and PCR-restriction fragment length polymorphism analysis of the spacer 16S-23S ribosomal DNA (rDNA); both methods were unable to differentiate R. paucula isolates. Eighteen strains belonging to other Ralstonia species (one R. eutropha strain, six R. pickettii strains, three R. solanacearum strains, and eight R. gilardii strains) were also tested by PCR-ribotyping, which failed to distinguish between the four species. The 16S-23S rDNA intergenic spacer of R. paucula contains the tRNA(Ile) and tRNA(Ala) genes, which are identical to genes described for R. pickettii and R. solanacearum.

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Figures

FIG. 1
FIG. 1
PCR-ribotyping patterns of Ralstonia species. (A) R. paucula strains. Lanes 1 to 4, type strains; lanes 5 and 6, Trousseau Hospital strains; lanes 7 to 13, other French hospital strains. (B) Ralstonia strains. Lane 1, R. paucula strain; lane 2, R. eutropha strain; lanes 3 to 8, R. pickettii strains; lanes 9 to 16, R. gilardii strains; lanes 17 to 19, R. solanacearum strains. Lanes M, molecular size markers; sizes are in base pairs.
FIG. 2
FIG. 2
Intergenic spacer region sequence of R. paucula (850 bp) (accession no. AF237657), with 153 bp belonging to the 16S portion (first set of boldface type), 521 bp corresponding to the intergenic spacer region, and the last 176 bp belonging to the 23S portion (last set of boldface type). The tRNAIle and tRNAAla genes (77 and 76 bp, respectively) are underlined in boldface.
FIG. 3
FIG. 3
Possible secondary structures of tRNAIle (left) and tRNAAla (right) of R. paucula with the three stem-loops and the anticodon region (shaded).
See this image and copyright information in PMC

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