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Multiplex polymerase chain reaction for detection of genes for Staphylococcus aureus thermonuclease and methicillin resistance and correlation with oxacillin resistance
Author(s) -
BRAKSTAD ODD G.,
MÆLAND JOHAN A.,
TVETEN YNGVAR
Publication year - 1993
Publication title -
apmis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0903-4641
DOI - 10.1111/j.1699-0463.1993.tb00165.x
Subject(s) - sccmec , coagulase , microbiology and biotechnology , staphylococcus aureus , multiplex polymerase chain reaction , penicillin binding proteins , polymerase chain reaction , penicillin , biology , gene , staphylococcus , methicillin resistant staphylococcus aureus , antibiotics , bacteria , genetics
A multiplex polymerase chain reaction (mPCR) was used for simultaneous amplification of the staphylococcal nuc gene, encoding the thermostable nuclease (TNase), and the mecA gene, encoding the penicillin‐binding protein 2a which is associated with staphyloccal methicillin resistance. A total of 219 staphylococcal strains were tested and the mPCR data were compared with coagulase production and in vitro oxacillin suscpetibility. The agreement was 100% for coagulase production and nuc amplification, and 97.7%, 96.8 and 97.3% for mecA amplication and oxacillin resistance tested with MIC determination, disk diffusion and agar screen methods, respectively. Discrepant results were due to non‐ S. aureus isolates with borderline MICs of oxacillin (1–8 μg/ml). In a pilot test the mPCR simultaneously amplified both genes of staphylococci in blood cultures. This mPCR is a rapid and reliable method for single‐step identification of cultures of MRSA and may prove to be useful for direct application on clinical specimens.