Identification of <I>Staphylococcus aureus</I> based on PCR amplification of species specific genomic 826 bp sequence derived from 44 kb SmaI-restriction fragment
| Authors | |
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| Year of publication | 2000 |
| Type | Article in Proceedings |
| Conference | 9th International Symposium on Staphylococci and Staphylococcal Infections |
| MU Faculty or unit | |
| Citation | |
| Field | Genetics and molecular biology |
| Description | The primers were prepared for PCR-amplification of a part genomic sequence specific to Staphylococcus aureus strains. They were derived from the 44 kb SmaI-restriction fragment of S. aureus genomic DNA (SmaI fragment L on the restriction map of strain NCTC 8325) common to all strains of this species (Pantucek et al. 1996, Int. J. Syst. Bacteriol. 46, 216-222). By hybridization of the labelled 44 kb SmaI-fragment to EcoRI-restriction patterns of the genomic DNA of 13 strains representative of S. aureus subsp. aureus the 2052 bp EcoRI-subfragment was proved to be common to all the tested strains. This fragment was used as the hybridization probe the specificity of which for the species S. aureus was tested in 41 species and subspecies of the genus Staphylococcus as well as in 7 strains of related bacterial genera. The probe hybridized only to the DNA of S. aureus subsp. aureus strains and S. aureus subsp. anaerobius. No hybridization was observed to the DNA of other staphylococcal as well as non-staphylococcal species. High specificity of the probe was verified in 203 culture collection and clinical S. aureus strains and no hybridization was observed in 45 collection and clinical coagulase-negative strains. The probe (i.e. genomic 2052 bp EcoRI-subfragment) was sequenced (accession number AJ132803) and used for designing PCR amplification primers, by means of which the amplification PCR-products of size 826 bp were obtained in all the tested S. aureus strains. The use of these primers enabled us to perform a rapid and reliable identification of strains belonging in the S. aureus subsp. aureus and to distinguish them from the other species of the genus Staphylococcus. |
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