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  • The aim of our work was to apply different molecular techniques to type MRSA strains, compare acquired results and performance of these methods. The collection of eight MRSA strains from different livestock farms in the Czech Republic was analysed. To type the strains, the MLST (Multi Locus Sequence typing), spa and SCCmec (Staphylococcal Chromosome Cassette mec) typing, rep-PCR analyse, ribotyping and PFGE (Pulsed-field gel electrophoresis) were applied. The (GTG)5 primer and the GelCompare software were used for the rep-PCR analyse. The ribotyping of the strains was performed on the RiboPrinterTM system, the SmaI restriction endonuclease was applied for the PFGE analyse. The SCCmec typing revealed presence of 4, 4D and 5 SCCmec type, the MLST analyse identified the ST8 and ST398 type among tested strains. Analysis of spa profiles revealed presence of five different strains. Also the rep-PCR indicated presence of five types of strains, however with distinct distribution. The ribotyping analysis seemed to be the most informative, as it identified six individual clone clusters. Unfortunately, the high cost and high technical requirements are still a drawback of this analysis. Surprisingly, the standard PFGE procedure was performed successfully only for two of the samples. The reason for this could be the presence of inhibitors in the samples, preventing the efficient PFGE - specific DNA isolation. The reproducibility of all investigated methods was considered as sufficient. With regard to the level of discrimination power, financial cost and equipment demands, we considered the rep-PCR as the most advantageous method for MRSA strain typing.
  • The aim of our work was to apply different molecular techniques to type MRSA strains, compare acquired results and performance of these methods. The collection of eight MRSA strains from different livestock farms in the Czech Republic was analysed. To type the strains, the MLST (Multi Locus Sequence typing), spa and SCCmec (Staphylococcal Chromosome Cassette mec) typing, rep-PCR analyse, ribotyping and PFGE (Pulsed-field gel electrophoresis) were applied. The (GTG)5 primer and the GelCompare software were used for the rep-PCR analyse. The ribotyping of the strains was performed on the RiboPrinterTM system, the SmaI restriction endonuclease was applied for the PFGE analyse. The SCCmec typing revealed presence of 4, 4D and 5 SCCmec type, the MLST analyse identified the ST8 and ST398 type among tested strains. Analysis of spa profiles revealed presence of five different strains. Also the rep-PCR indicated presence of five types of strains, however with distinct distribution. The ribotyping analysis seemed to be the most informative, as it identified six individual clone clusters. Unfortunately, the high cost and high technical requirements are still a drawback of this analysis. Surprisingly, the standard PFGE procedure was performed successfully only for two of the samples. The reason for this could be the presence of inhibitors in the samples, preventing the efficient PFGE - specific DNA isolation. The reproducibility of all investigated methods was considered as sufficient. With regard to the level of discrimination power, financial cost and equipment demands, we considered the rep-PCR as the most advantageous method for MRSA strain typing. (en)
Title
  • Typing of MRSA strains using different molecular-genetic methods
  • Typing of MRSA strains using different molecular-genetic methods (en)
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  • Typing of MRSA strains using different molecular-genetic methods
  • Typing of MRSA strains using different molecular-genetic methods (en)
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  • RIV/49608851:_____/11:#0000510!RIV12-MZE-49608851
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  • MRSA, typing, molecular techniques (en)
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  • Manga, Ivan
  • Vyletělová, Marcela
  • Karpíšková, R.
  • Šťástková, Z.
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