Successful control of the first OXA-48 and/or NDM carbapenemase-producing Klebsiella pneumoniae outbreak in Slovenia 2014-2016.

BACKGROUND: Carbapenemase-producing Enterobacteriaceae (CPE) occur only sporadically in Slovenia. AIM: To describe the first Slovenian carbapenemase-producing (CP) Klebsiella pneumoniae and Escherichia coli outbreak which occurred at the tertiary teaching hospital University Medical Centre Ljubljana from October 2014 to April 2015. METHODS: A CPE-positive case was defined as any patient infected or colonized with CPE. A strict definition of a contact patient was adopted. Measures to prevent cross-transmission included cohorting of all CPE carriers with strict contact precautions and assignment of dedicated healthcare workers, cohorting of all contact patients until obtaining the result of screening cultures, systematic rectal screening of contact patients, and tagging of all CPE-positive cases and their contacts. Educational campaigns on CPEs were implemented. Clinical specimens were processed using standard procedures. Pulsed-field gel electrophoresis (PFGE) was used to determine relatedness. Multi-locus sequence typing was performed on CP K. pneumoniae isolates that belonged to different pulsotypes. FINDINGS: Before the outbreak was brought under control, 40 patients were colonized or infected with OXA-48 and/or New Delhi metallo-ß-lactamase (NDM)-producing CPE; in 38 patients OXA-48 and/or NDM-producing K. pneumoniae was detected, in seven OXA-48 and/or NDM-producing E. coli was found together with K. pneumoniae, and in two patients only CP E. coli was isolated. The outbreak was oligoclonal with two major CP K. pneumoniae clusters belonging to ST437 and ST147 in epidemiologically linked patients. CONCLUSION: Initial standard control measures failed to prevent the outbreak. Once the problem had been recognized, strict infection control measures and the education of healthcare workers contributed to the successful control of the outbreak.

Survey of community-associated-methicillin-Resistant Staphylococcus aureus in Slovenia: identification of community-associated and livestock-associated clones.

The epidemiology of methicillin-resistant Staphylococcus aureus (MRSA) in Slovenia is poorly documented. The aim of this study was to investigate susceptibility patterns, virulence gene profile and clonality among MRSA isolates with positive screened resistance phenotype for CA-MRSA collected from patients in Slovenia, from January 2010 to December 2010. We included only MRSA isolates that were resistant to cefoxitin and oxacillin, and susceptible to at least two of the following four antibiotics: ciprofloxacin, erythromycin, clindamycin or gentamicin (presumptive CA-MRSA). Altogether 151 isolates fulfilled our screening phenotypic definition, 126 MRSA isolates were classified as CA-MRSA and 25 as HA-MRSA. Thirty-six per cent of them were resistant to ciprofloxacin, 24% to clindamycin, 33% to erythromycin and 13% to gentamicin. The mecA gene was detected in 150 isolates, while the mecC gene only in 1 isolate. The MRSA isolates were classified to 19 different clones. The most prevalent sequence types were ST5 (26.4%), ST45 (25.2%), ST22 (10.6%), ST398 (9.9%), ST8 (5.9%), ST7 (4.6%), ST1 (3.9%), ST152/377 (3.3%), ST228 (2.6%) and ST2883 (1.3%). The ST6, ST9, ST30, ST72, ST88, ST111, ST130, ST225 and ST772 were identified sporadically. The Panton-Valentine leukocidin (PVL) gene was detected in 13 (8.6%) isolates that belonged to ST5, ST7, ST8, ST22, ST72, ST88, ST 152/377 and ST772. Our results show high variability of CA-MRSA circulating in Slovenia and also the presence of LA-MRSA clones.

Detection of methicillin-resistant Staphylococcus aureus carrying the mecC gene in human samples in Slovenia.

SUMMARY Following the recognition of a mecC MRSA isolate from a patient hospitalized in the northeastern region of Slovenia, a national collection of 395 community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) isolates from 2006 to 2013 was screened. An additional six mecC MRSA strains were found and characterized as spa types t843, t9397 and t10009, and multilocus sequence type ST130. The low oxacillin minimum inhibitory concentrations and absence of the mecA gene make recognition of these MRSA strains problematical for diagnostic laboratories. In such strains the presence of mecC should be determined.

Intrathecal synthesis of specific antibodies in patients with invasion of the central nervous system by Mycoplasma pneumoniae.

Mycoplasma pneumoniae commonly causes respiratory tract infections in humans, but it may also be associated with central nervous system manifestations. The aim of the present study was to determine whether the cerebrospinal fluid taken from patients with neurologic symptoms due to Mycoplasma pneumoniae infection contains specific antibodies and whether the detection of these antibodies can be used for diagnosis. Mycoplasma pneumoniae was isolated from the cerebrospinal fluid taken from nine patients with central nervous system symptoms on admission to the hospital. In addition, Mycoplasma pneumoniae was detected in cerebrospinal fluid using polymerase chain reaction in four other patients. Antibodies to Mycoplasma pneumoniae were detected using the enzyme immunosorbent assay, indirect immunoperoxidase assay and immunoblotting in cerebrospinal fluid samples from 14 of 19 patients included in the study. The indirect immunoperoxidase assay showed high titers of Mycoplasma pneumoniae immunoglobulin G1 (IgG1) and IgM antibodies in cerebrospinal fluid samples of some patients with meningoencephalitis or meningitis. Titers of specific IgA, IgG2 and IgG3 antibodies were lower, while specific IgG4 was not detectable. Cerebrospinal fluid samples with higher antibody titers also contained IgA, IgG1, IgG2, IgG3 and IgM antibodies that recognized the P1 adhesin (170 kDa protein) of Mycoplasma pneumoniae. A comparison of antibody titers of concomitant serum/cerebrospinal fluid samples to Mycoplasma pneumoniae and those to measles virus by enzyme immunosorbent assay suggested the intrathecal synthesis of IgG and IgM antibodies to Mycoplasma pneumoniae in patients with acute meningoencephalitis. Data from this study clearly reinforce previous findings that Mycoplasma pneumoniae is an etiologic agent of central nervous system infections in humans.

Typing of consecutive methicillin-resistant Staphylococcus aureus isolates from intensive care unit patients and staff with pulsed-field gel electrophoresis.

Six consecutive methicillin-resistant Staphylococcus aureus (MRSA) isolates obtained in a 2-month period from tracheal aspirates of six intensive care unit (ICU) patients with nosocomial pneumonia and two MRSA isolates from nasal carriers among staff were typed to determine whether one or more strains were involved and whether nasal carriage was the source of the outbreak. Pulsed-field gel electrophoresis (PFGE) of chromosomal DNA was used to type MRSA isolates. The typing showed that the outbreak was caused by a single epidemic MRSA clone. The MRSA strain isolated from the staff was unrelated to the outbreak strain and was therefore not the source of the outbreak in this study. The source was apparently the index patient followed by transfer of MRSA to other patients on medical equipment or on the hands of staff who did not adhere strictly to infection control measures.