Characterization of Mycoplasmas by PCR and Sequence Analysis with Universal 16S rDNA Primers

Johansson, Karl‐Erik; Heldtander, M.; Pettersson, Bertil

doi:10.1385/0-89603-525-5:145

Cited by 62 publications

(64 citation statements)

References 28 publications

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“…These analyses were performed on 20 field strains following the standard procedures described by Johansson et al [17]. Briefly, in vitro amplification of the 16S rRNA genes was performed by seminested PCR, using the four universal primers (U1 (F), U8 (R), U2 (F), U5 (R)) described by Johansson et al [17], and was followed by sequence determination of the PCR products (Genome Express, Meylan, France).…”

Section: Characterization Of Mycoplasmas By Pcr and Sequence Analysismentioning

confidence: 99%

“…Briefly, in vitro amplification of the 16S rRNA genes was performed by seminested PCR, using the four universal primers (U1 (F), U8 (R), U2 (F), U5 (R)) described by Johansson et al [17], and was followed by sequence determination of the PCR products (Genome Express, Meylan, France). Evaluation of sequence data was performed on-line using a Basic Local Alignment Search Tool.…”

Section: Characterization Of Mycoplasmas By Pcr and Sequence Analysismentioning

confidence: 99%

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Assessment of PCR for routine identification of species of the Mycoplasma mycoides cluster in ruminants

et al. 2004

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-DNA amplification techniques offer considerable promise for the identification of Mycoplasma mycoides cluster members. They avoid antigenic cross-reactivity and variability that hamper serological methods. Many sets of primers, specific of these different members and of Mycoplasma putrefaciens, have been proposed. To assess the reliability of some of these PCR tests in routine laboratory diagnostic use, 230 field strains supposed to belong to this group were simultaneously identified by PCR and an antigenic method. The results were well correlated to antigenic identification for M. putrefaciens, but PCR failed to identify respectively 74% and 52% of M. mycoides subsp. mycoides Large Colony type and M. capricolum subsp. capricolum strains. Any identification of M. mycoides subsp. mycoides Small Colony type must be confirmed by two different tests. Difficulties in defining the M. species bovine serogroup 7 were also encountered with both the PCR and immunological methods. The occurrence of putative variable antigen(s) on the mycoplasma surface may explain part of the identification difficulties encountered with the immunological methods. Mycoplasma mycoides cluster / PCR / ruminants / identification / variable antigen

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Section: Characterization Of Mycoplasmas By Pcr and Sequence Analysismentioning

confidence: 99%

Section: Characterization Of Mycoplasmas By Pcr and Sequence Analysismentioning

confidence: 99%

Assessment of PCR for routine identification of species of the Mycoplasma mycoides cluster in ruminants

et al. 2004

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“…The Mollicutes split into two major branches: the AAP branch, containing the Acholeplasma, Anaeroplasma and Asteroleplasma genera, and the Candidatus Phytoplasma phyla; the other is the SEM branch that includes the Spiroplasma, Entomoplasma, Mesoplasma, Ureaplasma and Mycoplasma genera [Johansson et al, 1998;Maniloff, 1992;Razin et al, 1998]. Interestingly, the genus Mycoplasma is polyphyletic, with species clustering within the Spiroplasma, Pneumoniae and Hominis phylogenetic groups [Behbahani et al, 1993;Johansson et al, 1998;Maniloff, 1992]. Nevertheless, additional phylogenetic markers such as the elongation factor EF-Tu (tuf) gene, ribosomal protein genes, the 16S-23S rRNA intergenic sequences, etc, have been already used as complementary comparative data, thus there is no unique phylogenetic tree for Mollicutes [Razin et al, 1998].…”

Section: Pcr Sequencing Phylogeny and Molecular Epidemiologymentioning

confidence: 99%

Molecular Diagnostics of Mycoplasmas: Perspectives from the Microbiology Standpoint

Flores-Medina¹,

Soriano-Becerril²,

Díaz-García³

2012

Polymerase Chain Reaction

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“…If there are no other known mollicutes in the association, it is logical to proceed directly to cloning. Alternatively, preliminary identification can be by PCR and DNA sequencing using primers specific for eubacterial 16S rRNA genes (Johansson et al, 1998) or the 16S-23S ITS region (Maidak et al, 1997). If a simple BLAST search suggests that the 16S rRNA gene sequence of an isolate may be unique, a similarity matrix (Felsenstein, 1993) relating the candidate strain to its closest neighbours should be constructed.…”

Section: (Ii) Preliminary Identificationmentioning

confidence: 99%

Revised minimal standards for description of new species of the class Mollicutes (division Tenericutes)

Brown

Whitcomb

Bradbury

2007

International Journal of Systematic and Evolutionary Microbiology

138

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Minimal standards for novel species of the class Mollicutes (trivial term, mollicutes), last published in 1995, require revision. The International Committee on Systematics of Prokaryotes Subcommittee on the Taxonomy of Mollicutes proposes herein revised standards that reflect recent advances in molecular systematics and the species concept for prokaryotes. The mandatory requirements are: (i) deposition of the type strain into two recognized culture collections, preferably located in different countries; (ii) deposition of the 16S rRNA gene sequence into a public database, and a phylogenetic analysis of the relationships among the 16S rRNA gene sequences of the novel species and its neighbours; (iii) deposition of antiserum against the type strain into a recognized collection; (iv) demonstration, by using the combination of 16S rRNA gene sequence analyses, serological analyses and supplementary phenotypic data, that the type strain differs significantly from all previously named species; and (v) assignment to an order, a family and a genus in the class, with an appropriate specific epithet. The 16S rRNA gene sequence provides the primary basis for assignment to hierarchical rank, and may also constitute evidence of species novelty, but serological and supplementary phenotypic data must be presented to substantiate this. Serological methods have been documented to be congruent with DNA-DNA hybridization data and with 16S rRNA gene placements. The novel species must be tested serologically to the greatest extent that the investigators deem feasible against all neighbouring species whose 16S rRNA gene sequences show .0.94 similarity. The investigator is responsible for justifying which characters are most meaningful for assignment to the part of the mollicute phylogenetic tree in which a novel species is located, and for providing the means by which novel species can be identified by other investigators.

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Characterization of Mycoplasmas by PCR and Sequence Analysis with Universal 16S rDNA Primers

Cited by 62 publications

References 28 publications

Assessment of PCR for routine identification of species of the Mycoplasma mycoides cluster in ruminants

Assessment of PCR for routine identification of species of the Mycoplasma mycoides cluster in ruminants

Molecular Diagnostics of Mycoplasmas: Perspectives from the Microbiology Standpoint

Revised minimal standards for description of new species of the class Mollicutes (division Tenericutes)

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