Detection of metallo-ß-lactamase genes in clinical specimens by a commercial multiplex PCR system.

hyplex®-MBL ID Multiplex PCR-ELISA, a novel method for identifying metallo-ß-lactamase genes directly in clinical specimens, was evaluated using a consecutive collection of 326 samples from three hospitals in Greece characterized by high prevalence of VIM producers. The method exhibited high sensitivity (98.0%) and specificity (98.6%) and was proven reliable in detecting bla(VIM) genes in blood, urine, pus, and sputum samples that, as confirmed by conventional methods, contained various VIM-producing species. Future multicenter studies should be considered for the thorough evaluation of this method and its potential diagnostic utility.

High rates of community-acquired, Panton-Valentine leukocidin (PVL)- positive methicillin-resistant S. aureus (MRSA) infections in adult outpatients in Greece.

Staphylococcus aureus was isolated in 88 (30.8%) of 286 adult patients suffering from various skin and soft-tissue infections examined in the outpatient department of a 650 bed tertiary-care hospital of Athens, Greece between January 2006 and December 2007. Twenty-seven (30.7%) of the S. aureus infections were caused by methicillin-resistant S. aureus (MRSA). All MRSA isolates were also resistant to tetracycline, fucidic acid and kanamycin, but were sensitive to gentamicin and tobramycin, as well as to to cotrimoxazole, chloramphenicol, quinolones, clindamycin and erythromycin. All isolates belonged to staphylococcal cassette chromosome mec elements (SCCmec) type IV, and were found to carry the lukF-PV and lukS genes coding for Panton-Valentine leukocidin (PVL). Pulsed-field gel electrophoresis (PFGE) and spa-typing revealed high genetic similarity among all MRSA isolates and with the PFGE pattern of the well-described ST80 clone that seems to be spreading through Europe. The high prevalence of MRSA among S. aureus infections in the community signify that empiric therapy in Greece, when clinically indicated, should exclude beta-lactam antibiotics. Moreover, the establishment of an active screening for PVL-positive community-acquired (CA)-MRSA carriage and the adoption of a search and destroy strategy for CA-MRSA in all patients admitted with purulent skin and soft-tissue is of high priority in Greece as well as in all European countries which face high rates of CA-MRSA infection.

Acanthamoeba keratitis and contact lens disinfecting solutions.

OBJECTIVES: To report cases of culture-proved Acanthamoeba keratitis in Greece over a 10-year period and to evaluate the effectiveness of the commonly used commercial contact lens disinfecting systems in clinical cases of Acanthamoeba keratitis. MATERIAL AND METHODS: During the years 1994-2004, 45 contact lens wearers and 3 non-contact lens wearers presenting with symptoms and signs of keratitis underwent corneal sampling. The scrapings obtained were inoculated directly onto appropriate culture media for bacteria, fungi and Acanthamoeba. All proved positive for Acanthamoeba. The contact lenses and contact lens disinfecting solutions (16 one-step 3% hydrogen peroxide and 3 multipurpose solutions) of 19/45 patients with culture-proven Acanthamoeba keratitis were cultured for bacteria, fungi and Acanthamoeba. RESULTS: Acanthamoeba was isolated from contact lenses and contact lens disinfecting solutions in all 19 cases of Acanthamoeba keratitis studied. CONCLUSIONS: The main risk factor for corneal infection in contact lens wearers is the use of contact lens disinfecting systems ineffective at killing Acanthamoeba cysts and trophozoites, as well as bacteria and fungi. Improvement or development of new contact lens disinfecting systems by manufacturers is needed to prevent Acanthamoeba keratitis.