Mycobacterium arupense pulmonary infection: antibiotic resistance and restriction fragment length polymorphism analysis.

Mycobacterium arupense is a novel mycobacterium species. It was first identified from clinical specimens in 2006 and since then there have been only two reports of its recovery from clinical samples. In the present case M. arupense was isolated from the sputum of a 62-year-old man with a malignant mass in his left kidney, who presented with a one-month history of recurrent fever, dyspnea and haemoptysis. M. arupense was identified with sequencing of hsp65 and 16S rRNA genes. In the present study, its biochemical profile along with its resistance status and hsp65 RFLP analysis is presented.

Mycobacterium thermoresistibile: case report of a rarely isolated mycobacterium from Europe and review of literature.

Mycobacterium thermoresistibile is a non-tuberculous mycobacterium strongly associated with human infections. Since 1966, there have only been six reports of its isolation from clinical samples. We report on the first case from Europe and review all the previous cases. Identification was achieved with sequencing of the 16S rRNA and hsp65 genes. This study presents its phenotypic and biochemical profile, susceptibilities to selected antibiotics and hsp65 polymerase chain reaction-restriction fragment length polymorphism profile with BsteII and Hae III .

Identification of human papillomavirus DNA in melanoma biopsy specimens of Greek population.

The aim of this retrospective study was to evaluate melanoma biopsy specimens from the Greek population living in the prefecture of Larissa for the presence of human papillomavirus (HPV) DNA and to determine the possible relationship between HPV and clinical outcome in these patients. Twenty-eight melanoma biopsy specimens, 20 from primary cutaneous melanoma and eight from melanoma metastasis were obtained from 28 patients. The biopsy samples were formalin-fixed and paraffin wax-embedded. The control group consisted of three junctional melanocytic nevi, histologically confirmed, and three punch biopsies from normal skin that were obtained from six healthy individuals. The presence and types of HPV DNA were assessed by the amplification of a fragment of the LI region by consensus primer polymerase chain reaction (PCR) combined with restriction fragment length polymorphism analysis (RFLPA). In each biopsy specimen that was evaluated, HPV 6, HPV 11, HPV 16 and HPV 18 positive controls from genital HPV lesions were included. Five of 28 (17.85%) biopsy melanoma specimens were positive for HPV DNA. Conversely, HPV was not detected in any of the biopsy specimens of the control group (0/6). HPV viral type 16 was found in two samples and HPV 6 DNA in three. Our results regarding the possible relationship between melanoma and HPV DNA were not statistically significant (p > 0.05). These findings suggest that ultraviolet sun exposure remains the main cause of melanoma in our region. The role of cutaneous HPV infection in the pathogenesis of melanoma remains elusive.

Clonal dissemination of mupirocin-resistant staphylococci in Greek hospitals.

OBJECTIVES: To determine the rates of mupirocin resistance in staphylococci during a 4 year period (1999-2002) in Greece. MATERIALS: A total of 1200 Staphylococcus aureus and 2760 coagulase-negative staphylococci (CoNS), consecutively collected from four Greek hospitals located in different geographical areas, were tested for susceptibility to mupirocin using the Etest and a reference agar dilution method. RESULTS: Twenty-four S. aureus (2%) and 532 CoNS (19.2%) were found to be mupirocin-resistant during the study period. High-level mupirocin resistance was detected in 20 S. aureus (1.6%) and in 440 CoNS (15.9%), respectively. No variations in the rates of mupirocin-resistant S. aureus in relation to the year of collection were observed. In contrast, the rate of mupirocin-resistant CoNS increased dramatically from 9% in 1999, to 14% in 2000, 20% in 2001 and reached 33% in 2002. PFGE analysis revealed the presence of one main clone (A) among mupirocin-resistant S. aureus and two main clones (i and a) among Staphylococcus epidermidis isolates. CONCLUSIONS: In Greece, the rate of mupirocin-resistant S. aureus has remained low and steady since 1999. The high rate of mupirocin-resistant CoNS (33%) in 2002 was due mainly to clonal dissemination of epidemic hospital 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.

Multicenter evaluation of the fully automated bactec MGIT 960 system and three molecular methods for the isolation and the identification of mycobacteria from clinical specimens.

A combination of Bactec MGIT 960 system, a PCR-based assay and a PCR-Restriction Analysis procedure (PRA) was assessed for the detection and identification of mycobacteria from clinical samples. The MGIT recovered 243 from 266 mycobacterial isolates. The PCR-based assay correctly identified all (234) Mycobacterium tuberculosis isolates, while 35 nontuberculous mycobacterial isolates were identified by PRA.

Clinical isolates of macrolide-resistant Streptococcus pyogenes in Central Greece.

A total of 300 Streptococcus pyogenes isolates, collected during 2001 from five hospitals in the Thessalia district (Central Greece), were examined for their resistance to macrolides. Resistance to erythromycin was detected in 58 isolates (19.3%). Of these, 68.9% were susceptible to clindamycin (M-phenotype) and carried the mefA gene. Of the remaining isolates, 18 expressed the MLS(B) phenotype: 12 and six exhibited inducible and constitutive resistance to clindamycin, respectively. All of these strains were found to be ermA(TR) positive, except for four that had the ermB gene. Of the erythromycin-resistant strains, none was found to be resistant to penicillin, tetracycline or quinupristin-dalfopristin. Molecular typing by PFGE showed the presence of a limited number of clones.

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.

Dissemination of clonally unrelated erythromycin- and glycopeptide-resistant Enterococcus faecium isolates in a tertiary Greek hospital.

Between September 1999 to February 2001, 25 glycopeptide-resistant Enterococcus faecium (GRE) isolates were recovered from a Greek hospital. The isolates exhibited 13 distinct chromosomal macrorestriction types by pulsed-field gel electrophoresis, and all were erythromycin and vancomycin resistant, carrying the genes vanA and ermB. Vancomycin resistance, always linked with erythromycin resistance, was transferable from 17 isolates. The dissemination of erythromycin-resistant GRE strains may, at least in part, reflect the extensive use of macrolides in husbandry in Greece.

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.