Preparation of a speciesspecific molecular probe for identification of Staphylococcus aureus
| Autoři | |
|---|---|
| Rok publikování | 2000 |
| Druh | Článek v odborném periodiku |
| Časopis / Zdroj | Scripta medica |
| Fakulta / Pracoviště MU | |
| Citace | |
| Obor | Genetika a molekulární biologie |
| Popis | In our previous work a 44-kb genomic SmaI-restriction fragment common to all S. aureus strains has been described. This restriction fragment was used as a starting point in preparation of a species-specific hybridisation probe for rapid molecular identification S. aureus strains. EcoRI-restricted genomic DNAs isolated from 13 different S. aureus strains, including the type strain S. aureus CCM 885, and from the type strain S. epidermidis CCM 2124 (negative control) were hybridised to a probe prepared from the whole sequence of the 44-kb fragment. The EcoRI-restriction fragments showing strong hybridisation to the S. aureus 44-kb probe were cloned in the pUCl8 vector in E. coli and used as candidates for the preparation of molecular probes specific for S. aureus. The cloned 2-kb EcoRI-restriction fragment was chosen as the most specific fragment that hybridised to the homologic EcoRl-fragment present in genomic DNA of more than 100 S. aureus strains of different provenance. No hybridisation was detected on genomic DNA isolated from S. epidennidis and the other species of the genus Sraphylococcus. The 2-kb EcoRl-fragment was sequenced and the primers for rapid PCR-identification of S. aureus strains were derived. In using these primers, 825 b PCR amplification products were obtained in DNA isolated from 100 S. aureus strains. No PCR amplification products were obtained in DNA isolated from the other Staphylococcus species. |
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