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Evaluation of tRNA gene PCR for identification of Mollicutes

Tim Stakenborg, Jo Vicca, Rita Verhelst UGent, Patrick Butaye UGent, Dominiek Maes UGent, Anne Naessens, Geert Claeys UGent, CATHARINE DE GANCK UGent, Freddy Haesebrouck UGent and Mario Vaneechoutte UGent (2005) JOURNAL OF CLINICAL MICROBIOLOGY. 43(9). p.4558-4566
abstract
We evaluated the applicability of IRNA 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 IRNA 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.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
POLYMERASE CHAIN-REACTION, LENGTH POLYMORPHISM ANALYSIS, INTERGENIC SPACER PCR, UREAPLASMA-UREALYTICUM, MYCOPLASMA-GENITALIUM, ACHOLEPLASMA-LAIDLAWII, MONOCLONAL-ANTIBODIES, TDNA-PCR, STRAINS, DNA
journal title
JOURNAL OF CLINICAL MICROBIOLOGY
J. Clin. Microbiol.
volume
43
issue
9
pages
4558-4566 pages
Web of Science type
Article
Web of Science id
000232020400038
JCR category
MICROBIOLOGY
JCR impact factor
3.537 (2005)
JCR rank
20/85 (2005)
JCR quartile
1 (2005)
ISSN
0095-1137
DOI
10.1128/JCM.43.9.4558-4566.2005
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
324515
handle
http://hdl.handle.net/1854/LU-324515
date created
2006-01-06 15:50:00
date last changed
2010-06-23 11:35:12
@article{324515,
  abstract     = {We evaluated the applicability of IRNA 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 IRNA 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.},
  author       = {Stakenborg, Tim and Vicca, Jo and Verhelst, Rita and Butaye, Patrick and Maes, Dominiek and Naessens, Anne and Claeys, Geert and DE GANCK, CATHARINE and Haesebrouck, Freddy and Vaneechoutte, Mario},
  issn         = {0095-1137},
  journal      = {JOURNAL OF CLINICAL MICROBIOLOGY},
  keyword      = {POLYMERASE CHAIN-REACTION,LENGTH POLYMORPHISM ANALYSIS,INTERGENIC SPACER PCR,UREAPLASMA-UREALYTICUM,MYCOPLASMA-GENITALIUM,ACHOLEPLASMA-LAIDLAWII,MONOCLONAL-ANTIBODIES,TDNA-PCR,STRAINS,DNA},
  language     = {eng},
  number       = {9},
  pages        = {4558--4566},
  title        = {Evaluation of tRNA gene PCR for identification of Mollicutes},
  url          = {http://dx.doi.org/10.1128/JCM.43.9.4558-4566.2005},
  volume       = {43},
  year         = {2005},
}

Chicago
Stakenborg, Tim, Jo Vicca, Rita Verhelst, Patrick Butaye, Dominiek Maes, Anne Naessens, Geert Claeys, CATHARINE DE GANCK, Freddy Haesebrouck, and Mario Vaneechoutte. 2005. “Evaluation of tRNA Gene PCR for Identification of Mollicutes.” Journal of Clinical Microbiology 43 (9): 4558–4566.
APA
Stakenborg, T., Vicca, J., Verhelst, R., Butaye, P., Maes, D., Naessens, A., Claeys, G., et al. (2005). Evaluation of tRNA gene PCR for identification of Mollicutes. JOURNAL OF CLINICAL MICROBIOLOGY, 43(9), 4558–4566.
Vancouver
1.
Stakenborg T, Vicca J, Verhelst R, Butaye P, Maes D, Naessens A, et al. Evaluation of tRNA gene PCR for identification of Mollicutes. JOURNAL OF CLINICAL MICROBIOLOGY. 2005;43(9):4558–66.
MLA
Stakenborg, Tim, Jo Vicca, Rita Verhelst, et al. “Evaluation of tRNA Gene PCR for Identification of Mollicutes.” JOURNAL OF CLINICAL MICROBIOLOGY 43.9 (2005): 4558–4566. Print.