Typing of Staphylococcus aureus and Staphylococcus epidermidis strains by PCR analysis of inter-IS256 spacer length polymorphisms.
IS256 elements are present in multiple copies in the staphylococcal genome, either flanking the transposon Tn4001 or independent of it. PCR-based analysis of inter-IS256 spacer polymorphisms was developed for typing of methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus epidermidis strains. Using SmaI macrorestriction analysis resolved by pulsed-field gel electrophoresis (PFGE) as the reference method for MRSA typing, excellent reproducibility (100%), discriminatory power (97%), and in vivo stability were observed. Good concordance of the results with those of other molecular typing methods was found for two MRSA collections. Inter-IS256 PCR analysis of a U.S. collection of MRSA strains (n = 36), previously characterized by 15 typing methods, showed more limited discrimination. Agreement was 78% with PFGE analysis and 83% with ribotyping (HindIII). Analysis of a second set of Belgian MRSA strains (n = 17), categorized into two widespread epidemic clones by PFGE analysis, showed 65% agreement. For typing of S. epidermidis strains (n = 26), inter-IS256 PCR showed complete typeability (100%) and good discriminatory power (85%). Inter-IS256 PCR analysis is proposed as an efficient molecular typing assay for epidemiological studies of MRSA or S. epidermidis isolates.
Deplano
A
A
Department of Microbiology, Hôpital Erasme, School of Public Health, Université Libre de Bruxelles, Brussels, Belgium.
Vaneechoutte
M
M
Verschraegen
G
G
Struelens
M J
MJ
D015373:Bacterial Typing Techniques; D001530:Belgium / Q000453:epidemiology; D017931:DNA Primers; D004251:DNA Transposable Elements / Q000235:genetics; D015252:Deoxyribonucleases, Type II Site-Specific; D016521:Electrophoresis, Gel, Pulsed-Field; D005069:Evaluation Studies as Topic; D016680:Genome, Bacterial; D006801:Humans; D016106:Methicillin Resistance / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000453:epidemiology; D013211:Staphylococcus aureus / Q000145:classification* / Q000187:drug effects / Q000235:genetics; D013212:Staphylococcus epidermidis / Q000145:classification* / Q000187:drug effects / Q000235:genetics
D017931:DNA Primers; D004251:DNA Transposable Elements; C025651:CCCGGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific