Identification of Staphylococcus aureus based on PCR amplification of species specific genomic 826 bp sequence derived from a common 44-kb Sma I restriction fragment

• Published in: Molecular and cellular probes Vol. 15; no. 5; pp. 249 - 257
• Main Authors: Štěpán, J., Pantůček, R., Růžičková, V., Rosypal, S., Hájek, V., Doškař, J.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.10.2001
• Subjects: Deoxyribonucleases, Type II Site-Specific - metabolism; DNA Primers; DNA Probes; Genome, Bacterial; Humans; Nucleic Acid Hybridization; Polymerase Chain Reaction; Polymorphism, Restriction Fragment Length; Sensitivity and Specificity; Staphylococcus aureus - classification; Staphylococcus aureus - genetics; Staphylococcus aureus, polymerase chain reaction, species-specific probe, 44-kb Sma I fragment L of NCTC 8325, molecular identification; Staphylococcus aureus, polymerase chain reaction, species-specific probe, 44-kb Sma I fragment L of NCTC 8325, molecular identification
• ISSN: 0890-8508; 1096-1194
• DOI: 10.1006/mcpr.2001.0368

Primers were designed for polymerase chain reaction (PCR)-amplification of a genomic sequence specific to Staphylococcus aureus strains. The sequence corresponds to a part of the 44-kb Sma I fragment (fragment L on the S. aureus NCTC 8325 restriction map) which was found to be common to strains of the S. aureus species (Pantůček et al 1996, International Journal of Systematic Bacteriology, 46: 216–222). The labelled 44-kb Sma I restriction fragment derived from S. aureus NCTC 8325–4 was hybridized to theEco RI restriction patterns of genomic DNA from 13 strains representing different macrorestriction types of S. aureus subsp. aureus. This made it possible to reveal the 2052 bp Eco RI restriction subfragment and to demonstrate its presence in all the tested strains. From the sequence of this subfragment, primers were designed by means of which the 826 bp amplicons were obtained in all 216 tested strains of S. aureus. No hybridization and PCR-products were observed in 40 collection strains of other staphylococcal species and subspecies as well as in 45 clinical strains of coagulase-negative staphylococci. These results lead us to the conclusion that the use of the above primers makes it possible to identify rapidly and reliably S. aureus strains of various provenance and different genotypes.