Detecting methicillin-resistant Staphylococcus epidermidis--disc diffusion, broth breakpoint or polymerase chain reaction?
Growth conditions are important for the expression of resistance to methicillin among staphylococci. Consequently a phenotypic susceptibility test has to be chosen carefully to avoid false susceptible results. In this study we wanted to devise rapid and simple phenotypic tests whose results completely correlate with the presence of the methicillin resistance gene, mecA. A simplified polymerase chain reaction (PCR) method not needing separate DNA extraction from the tested bacteria was used to amplify a 449 bp region of the mecA gene. One hundred and ten strains of S. epidermidis were tested. The results were in complete agreement with those from a broth tube breakpoint test, known to identify more strains as resistant than does the method recommended by NCCLS. In disc diffusion test it was possible to clearly distinguish resistant from susceptible strains by using discs containing oxacillin, cephalexin and cephradine. A 5 micrograms cephradine disc was further analysed by testing another 441 consecutive clinical isolates of staphylococci. All resistant coagulase-negative staphylococci grew out to the edge of this disc, whereas susceptible strains showed an inhibition zone at least 10 mm in diameter. The 5 micrograms cephradine disc is recommended for routine work. The PCR method and broth tube breakpoint test are both reliable reference methods.
Hedin
G
G
Department of Clinical Microbiology, University Hospital, Uppsala, Sweden.
Löfdahl
S
S
D001483:Base Sequence; D002506:Cephalexin / Q000633:toxicity*; D002515:Cephradine / Q000633:toxicity*; D005798:Genes, Bacterial; D016106:Methicillin Resistance / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010068:Oxacillin / Q000633:toxicity*; D016133:Polymerase Chain Reaction; D013212:Staphylococcus epidermidis / Q000187:drug effects / Q000235:genetics / Q000254:growth & development*
D015345:Oligonucleotide Probes; D002515:Cephradine; D002506:Cephalexin; D010068:Oxacillin