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

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 Smal fragment (fragment L on the S. aure us NCTC 8325 restriction map) which was found to be common to strains of th e S. aureus species (Pantucek et al., 1996, International journal of System atic Bacteriology, 46: 216-222). The labelled 44-kb Smal restriction fragme nt derived from S. aureus NCTC 8325-4 was hybridized to the EcoRI restricti on patterns of genomic DNA from 13 strains representing different macrorest riction types of S. aureus subsp. aureus. This made it possible to reveal t he 2052 bp EcoRI restriction subfragment and to demonstrate its presence in all the tested strains. From the sequence of this subfragment, primers wer e 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 observe d in 40 collection strains of other staphylococcal species and subspecies a s 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 p rovenance and different genotypes. (C) 2001 Academic Press.