Evaluation of tRNA gene PCR for identification of mollicutes

Research output: Journal Publications and ReviewsRGC 21 - Publication in refereed journalpeer-review

13 Scopus Citations
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Author(s)

  • Tim Stakenborg
  • Jo Vicca
  • Rita Verhelst
  • Dominiek Maes
  • Anne Naessens
  • Geert Claeys
  • Catharine De Ganck
  • Freddy Haesebrouck
  • Mario Vaneechoutte

Detail(s)

Original languageEnglish
Pages (from-to)4558-4566
Journal / PublicationJournal of Clinical Microbiology
Volume43
Issue number9
Publication statusPublished - Sept 2005
Externally publishedYes

Abstract

We evaluated the applicability of tRNA gene PCR in combination with fluorescent capillary electrophoresis with an ABI310 genetic analyzer (Applied Biosystems, Calif.) for the identification of different mollicute species. A total of 103 strains and DNA extracts of 30 different species belonging to the genera Acholeplasma, Mycoplasma, and Ureaplasma were studied. Reproducible peak profiles were generated for all samples, except for one M. genitalium isolate, the three M. gallisepticum isolates, and 8 of the 24 Ureaplasma cultures, where no amplification could be obtained. Clustering revealed numerous discrepancies compared to the identifications that had been previously obtained by means of biochemical and serological tests. Final identification was obtained by 16S rRNA gene amplification followed by sequence analysis and/or restriction digestion. This confirmed the identification obtained by tRNA gene PCR in all cases. Seven samples yielded an unexpected tRNA gene PCR profile. Sequence analysis of the 16S rRNA genes showed that six of these samples were mixed and that one had a unique sequence that did not match any of the published sequences, pointing to the existence of a not-yet-described species. In conclusion, we found tRNA gene PCR to be a rapid and discriminatory method to correctly identify a large collection of different species of the class of Mollicutes and to recognize not-yet-described groups. Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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Citation Format(s)

Evaluation of tRNA gene PCR for identification of mollicutes. / Stakenborg, Tim; Vicca, Jo; Verhelst, Rita et al.
In: Journal of Clinical Microbiology, Vol. 43, No. 9, 09.2005, p. 4558-4566.

Research output: Journal Publications and ReviewsRGC 21 - Publication in refereed journalpeer-review