M. Maniati

Novel Variant (blaVIM-4) of the Metallo-β-Lactamase Gene blaVIM-1 in a Clinical Strain of Pseudomonas aeruginosa

ABSTRACT A Pseudomonas aeruginosa isolate highly resistant to carbapenems was collected from a patient with postsurgical cerebrospinal infection in Greece. The isolate carried a class 1 integron that contained as a sole cassette the gene blaVIM-4, a novel variant of blaVIM-1, with one nucleotide difference resulting in a Ser-to-Arg change at amino acid position 175 of the VIM-1 enzyme. This is the first detection of a VIM-1 variant after its appearance in Italy.

Survey of methicillin-resistant coagulase-negative staphylococci in the hospitals of central Greece

A sample of 450 consecutive, non-replicated coagulase-negative staphylococci (CoNS), collected from clinical specimens during the period 2000-2001 from the five major hospitals of Thessaly district (Central Greece) were investigated for resistance to methicillin. Most of the isolates had been collected in a sporadic fashion from the intensive care units and the surgical wards of the participating hospitals. The majority of the isolates (76%) were Staphylococcus epidermidis (50%), Staphylococcus haemolyticus (14.8%) and Staphylococcus hominis (11.1%). All 316 isolates (70%) were classified as resistant according to NCCLS breakpoints (MIC > or =0.5 mg/l); 268 (59.5%) of them were mecA-positive in a PCR-based assay. All isolates with MIC > or =8 mg/l carried the gene, while, only 23.8% of isolates with MIC, 0.5-4 mg/l were carriers. Only 9% of the mecA-positive isolates were found to be sensitive to various non-beta-lactams, while 41.8% of the isolates were resistant to more than three antimicrobial groups apart from beta-lactams. Molecular typing by PFGE showed apparent heterogeneity among isolates of each species and the absence of predominant clones.

Evaluation of a novel method based on PCR Restriction Fragment Length Polymorphism Analysis of the tuf gene for the identification of Staphylococcus species.

A novel method, based on PCR Restriction Fragment Length Polymorphism Analysis (PRA) of a part of the tuf gene (370 bp), was designed for the identification of 11 staphylococcal species, including the most common staphylococcal pathogens. A total of 258 clinical isolates were validated by this assay, and the results were in concordance with those obtained by the reference method of Kloos and Schleifer.

Methicillin-resistant Staphylococcus aureus in the hospitals of Central Greece

A total of 250 consecutive Staphylococcus aureus clinical isolates were collected during the period 1999-2000 from the five major hospitals of the district of Thessaly (Central Greece). Thirty seven (14.8%) of the isolates were mecA-positive (MRSA) in a PCR-based assay; all exhibited resistance to oxacillin (agar dilution MICs > or =4 mg/L) and were also resistant to multiple antibiotics. Most of the MRSA isolates had been collected in the intensive care units and the surgical wards of the participating hospitals in a sporadic fashion. The MRSA incidence found here was significantly lower than reported in previous studies from Greece. Molecular typing by PFGE showed that the MRSA isolates were distributed between three pulsotypes. Evaluation of various conventional methods for assessing methicillin resistance showed that oxacillin agar dilution and immunological detection of PBP2a with the Slidex MRSA Detection kit were the most reliable in this setting. Misclassifications of isolates exhibiting low-level resistance (oxacillin MIC 2-4 mg/L) occurred with the salt agar screen, the oxacillin disk diffusion and the ATB Staph System methods.

Multicenter evaluation of the fully automated Bactec MGIT 960 system for susceptibility testing of Mycobacterium tuberculosis to pyrazinamide: comparison with the radiometric Bactec 460TB system.

One hundred and fifty clinical isolates of Mycobacterium tuberculosis were tested for susceptibility to pyrazinamide using the fully automated Bactec MGIT 960 system and the radiometric Bactec 460TB system. The overall concordance rate between MGIT 960 and radiometric system was 100% and the mean turnaround times to report the susceptibility test results were almost identical (6.37 and 6.8 days, respectively).