Extended Bibliography: |
Show bibliography
Ref #: |
14410 |
Author(s): |
Kong,F.;James,G.;Gordon,S.;Zelynski,A.;Gilbert,G.L. |
Journal: |
Appl Environ Microbiol |
Title: |
Species-specific PCR for identification of common contaminant mollicutes in cell culture |
Volume: |
67 |
Page(s): |
3195-200 |
Year: |
2001 |
Keyword(s): |
GENBANK/AF294989
GENBANK/AF294990
GENBANK/AF294991
GENBANK/AF294992
GENBANK/AF294993
GENBANK/AF294994
GENBANK/AF294995
GENBANK/AF294996
Cell Line/*microbiology
DNA Primers
DNA, Ribosomal Spacer/genetics
Humans
Molecular Sequence Data
Mollicutes/*classification/genetics/*isolation & purification
Polymerase Chain Reaction/*methods
RNA, Ribosomal, 16S/genetics
RNA, Ribosomal, 23S/genetics
Sensitivity and Specificity
Sequence Analysis, DNA
Species Specificity
|
Remarks: |
Mycoplasma arginini, M. fermentans, M. hyorhinis, M. orale, and Acholeplasma laidlawii are the members of the class Mollicutes most commonly found in contaminated cell cultures. Previous studies have shown that the published PCR primer pairs designed to detect mollicutes in cell cultures are not entirely specific. The 16S rRNA gene, the 16S-23S rRNA intergenic spacer region, and the 5' end of the 23S rRNA gene, as a whole, are promising targets for design of mollicute species-specific primer pairs. We analyzed the 16S rRNA genes, the 16S-23S rRNA intergenic spacer regions, and the 5' end of the 23S rRNA genes of these mollicutes and developed PCR methods for species identification based on these regions. Using high melting temperatures, we developed a rapid-cycle PCR for detection and identification of contaminant mollicutes. Previously published, putative mollicute-specific primers amplified DNA from 73 contaminated cell lines, but the presence of mollicutes was confirmed by species-specific PCR in only 60. Sequences of the remaining 13 amplicons were identified as those of gram-positive bacterial species. Species-specific PCR primers are needed to confirm the presence of mollicutes in specimens and for identification, if required. |
URL: |
11425741 |
|
Ref #: |
95481 |
Author(s): |
Kim,K.S.;Ko,K.S.;Chang,M.W.;Hahn,T.W.;Hong,S.K.;Kook,Y.H. |
Journal: |
FEMS Microbiol Lett |
Title: |
Use of rpoB sequences for phylogenetic study of Mycoplasma species |
Volume: |
226 |
Page(s): |
299-305 |
Year: |
2003 |
Keyword(s): |
GENBANK/AY191418
GENBANK/AY191419
GENBANK/AY191420
GENBANK/AY191421
GENBANK/AY191422
GENBANK/AY191423
GENBANK/AY191424
GENBANK/AY191425
GENBANK/AY191426
GENBANK/AY191427
GENBANK/AY191428
GENBANK/AY191429
GENBANK/AY191430
GENBANK/AY191431
GENBANK/AY191432
GENBANK/AY191433
GENBANK/AY191434
GENBANK/AY191435
GENBANK/AY191436
GENBANK/AY191437
GENBANK/AY191438
GENBANK/AY191439
GENBANK/AY191440
GENBANK/AY191441
GENBANK/AY191442
GENBANK/AY191443
Amino Acid Sequence
DNA, Ribosomal/chemistry/genetics
DNA-Directed RNA Polymerases/*chemistry/*genetics
Genes, Bacterial
Molecular Sequence Data
Mycoplasma/classification/*genetics
*Phylogeny
RNA, Ribosomal, 16S/genetics
Sequence Alignment
Sequence Homology, Amino Acid
Sequence Homology, Nucleic Acid
Ureaplasma/genetics
|
Remarks: |
rpoB sequences encoding the beta-subunit of RNA polymerase were determined in 26 Mycoplasma species for phylogenetic study. The portion of rpoB DNA used in this study showed a high degree of variation in terms of size and sequence among species. The rpoB phylogenies inferred from amino acid and nucleotide sequences were used to divide the mycoplasmas into two groups, a 'pneumoniae group' and a 'hominis group', which was consistent with the result from 16S rDNA sequence analysis. However, phylogenetic relationships within these groups differed in the two gene trees, which were supported by the incongruence length difference (ILD) test. This indicates that multiple gene sequences must be applied to infer accurate phylogenetic relationships among the mycoplasmas. The rpoB sequence, and especially the deduced amino acid sequence, offers a good alternative marker. |
URL: |
14553926 |
|
Ref #: |
11947 |
Author(s): |
Kong,F.;James,G.;Gordon,S.;Zelynski,A.;Gilbert,G.L. |
Journal: |
Appl Environ Microbiol |
Title: |
Species-specific PCR for identification of common contaminant mollicutes in cell culture |
Volume: |
67 |
Page(s): |
3195-200 |
Year: |
2001 |
Keyword(s): |
GENBANK/AF294989
GENBANK/AF294990
GENBANK/AF294991
GENBANK/AF294992
GENBANK/AF294993
GENBANK/AF294994
GENBANK/AF294995
GENBANK/AF294996
Cell Line/*microbiology
DNA Primers
DNA, Ribosomal Spacer/genetics
Human
Molecular Sequence Data
Mollicutes/*classification/genetics/*isolation & purification
Polymerase Chain Reaction/*methods
RNA, Ribosomal, 16S/genetics
RNA, Ribosomal, 23S/genetics
Sensitivity and Specificity
Sequence Analysis, DNA
Species Specificity
|
Remarks: |
Mycoplasma arginini, M. fermentans, M. hyorhinis, M. orale, and Acholeplasma laidlawii are the members of the class Mollicutes most commonly found in contaminated cell cultures. Previous studies have shown that the published PCR primer pairs designed to detect mollicutes in cell cultures are not entirely specific. The 16S rRNA gene, the 16S-23S rRNA intergenic spacer region, and the 5' end of the 23S rRNA gene, as a whole, are promising targets for design of mollicute species-specific primer pairs. We analyzed the 16S rRNA genes, the 16S-23S rRNA intergenic spacer regions, and the 5' end of the 23S rRNA genes of these mollicutes and developed PCR methods for species identification based on these regions. Using high melting temperatures, we developed a rapid-cycle PCR for detection and identification of contaminant mollicutes. Previously published, putative mollicute-specific primers amplified DNA from 73 contaminated cell lines, but the presence of mollicutes was confirmed by species-specific PCR in only 60. Sequences of the remaining 13 amplicons were identified as those of gram-positive bacterial species. Species-specific PCR primers are needed to confirm the presence of mollicutes in specimens and for identification, if required. |
URL: |
21318714 |
|
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