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Genomic Variability of Staphylococcus aureus and the Other Coagulase-Positive Staphylococcus Species Estimated by Macrorestriction Analysis Using Pulsed-Field Gel Electrophoresis

ROMAN PANTEK¹, FRIEDRICH GÖTZ², JIÍ DOKA¹ and STANISLAV ROSYPAL^1,*

¹ Department of Genetics and Molecular Biology, Faculty of Science, Masaryk University, 611 37 Brno, Czech Republic
² Lehrstuhl für Mikrobielle Genetik, Universität Tübingen, 72076 Tübingen 1, Germany

^* Corresponding author. Mailing address: Department of Genetics and Molecular Biology, Faculty of Science, Masaryk University, Kotláská 2, 611 37 Brno, Czech Republic. Phone: (0042-5)4112 9549. Fax: (0042-5)4121 1214. Electronic mail address: rosypal{at}sci.muni.cz.

ABSTRACT

The genomic DNAs of 95 culture collection and hospital Staphylococcus aureus subsp. aureus strains of various origins, as well as the genomic DNAs of other coagulase-positive Staphylococcus species, were cleaved with restriction endonuclease Sma I and subjected to pulsed-field gel electrophoresis. The levels of similarity of the Sma I restriction patterns of the S. aureus subsp. aureus strains varied from 30 to 100%, which is considered characteristic of this species; thus, these organisms belonged to the same species restriction group. Within this range of similarity values 13 S. aureus intraspecies restriction groups were identified, and each group consisted of strains whose levels of similarity ranged from 65 to 100%. S. aureus subsp. aureus CCM 885^T (T = type strain) belonged to the major intraspecies restriction group that comprised 39% of the S. aureus strains which we studied. The strains of the other coagulase-positive staphylococci, including Staphylococcus aureus subsp. anaerobius, Staphylococcus hyicus, Staphylococcus intermedius, Staphylococcus delphini, and Staphylococcus schleiferi subsp. coagulans, clustered with their type strains in separate restriction groups. S. aureus subsp. aureus exhibited almost no similarity to these species. We found 44-kb Sma I fragments in all of the S. aureus subsp. aureus and S. aureus subsp. anaerobius strains studied, and these fragments are considered characteristic of the species S. aureus. The high level of homology of these fragments was confirmed by the results of DNA hybridization experiments in which we used representatives of individual intraspecies restriction groups. Of the other staphylococci studied, only Staphylococcus epidermidis and one strain of S. hyicus contained these fragments. However, the levels of homology between these fragments and the fragments of S. aureus were found to be very low.

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