PCR-Based assay for discrimination between invasive and contaminating Staphylococcus epidermidis strains.

The discrimination between Staphylococcus epidermidis strains that contaminate and infect blood cultures is a daily challenge for clinical laboratories. The results of PCR detection of putative virulence genes were compared for contaminating strains, sepsis-related strains, catheter strains, and saprophytic strains. Multiplex PCR was used to explore the atlE gene, which is involved in initial adherence, the intercellular adhesion gene cluster (ica), which mediates the formation of the biofilm, and the agrA, sarA, and mecA genes, which might contribute to the pathogenicity of S. epidermidis. Whereas the atlE, agrA, and sarA genes were almost ubiquitously amplified, the ica and mecA genes were detected significantly more in infecting strains than in contaminating strains (P

Evidence for nasal carriage of methicillin-resistant staphylococci colonizing intravascular devices.

Nasal surveillance cultures were performed for 54 patients exhibiting >/=10(3) CFU of methicillin-resistant coagulase-negative staphylococci per ml in central venous catheter (CVC) rinse cultures over a 6-month period. Forty-two of the nasal cultures yielded growth of methicillin-resistant coagulase-negative staphylococci, and 33 of the 42 cultures contained organisms that belonged to the same species as the CVC isolates. Of the 33 same-species isolates, 20 appeared to be identical strains by pulsed-field gel electrophoresis analysis. These data suggest that measures should be taken to reduce cross-contamination between the respiratory tract and intravascular devices. However, the potential interest in detecting methicillin-resistant coagulase-negative staphylococcus carriage in high-risk patients is hampered by the lack of sensitivity of nasal surveillance cultures.

Comparison of ATB staph, rapid ATB staph, Vitek, and E-test methods for detection of oxacillin heteroresistance in staphylococci possessing mecA.

The performance characteristics of the E-test (AB Biodisk, Solna, Sweden), the ATB Staph, the Rapid ATB Staph, and the Vitek GPS-503 card (bioMérieux, La Balme Les Grottes, France) methods for the detection of oxacillin resistance in a collection of staphylococci with a high proportion of troublesome strains were evaluated. Sixty-four Staphylococcus aureus strains and 76 coagulase-negative staphylococcal strains were tested. All strains were mecA positive and were characterized by the oxacillin agar screen plate test; 75 (53.6%) were found to be heterogeneous by a large-inoculum oxacillin disk diffusion assay, and oxacillin MICs for 89 (63.6%) were < or = 32 microg/ml. Three (4.7%) S. aureus strains and 25 (32.9%) coagulase-negative strains were classified as susceptible by the E-test, as defined by the National Committee for Clinical Laboratory Standards (NCCLS) oxacillin breakpoint (MIC < or = 2 microg/ml). The ATB Staph method failed to detect oxacillin resistance in 7 (11%) S. aureus isolates and 32 (42.1%) coagulase-negative isolates. The MICs for all but six of these discrepant isolates were < or = 16 microg/ml. The Rapid ATB Staph method was tested against S. aureus strains only and yielded 15 (23.4%) false-susceptible results for strains for which the MICs were < or = 32 microg/ml. The Vitek system was the best-performing system, since it failed to detect oxacillin resistance in only 3 (4.7%) S. aureus strains and 15 (19.7%) coagulase-negative strains, the MICs for all of which were < or = 2 microg/ml. These data indicate that (i) the performance of the two ATB Staph systems can be limited when the prevalence of borderline-heteroresistant staphylococci is high and (ii) the unreliability of the E-test and the Vitek methods for detecting resistant coagulase-negative strains might be reduced by the potential revision of the oxacillin breakpoint currently recommended by the NCCLS.