BACKGROUND: This study presents the results of a multidisciplinary, nosocomial MRSA outbreak investigation in an 8-bed medical intensive care unit (ICU). The identification of seven MRSA positive patients in the beginning of 2014 led to the closure of the ward for several weeks. A multidisciplinary, retrospective investigation was initiated in order to identify the reason and the source for the outbreak, describe MRSA transmission in the department and identify limitations in infection control. METHODS: The investigation comprised an epidemiological description of MRSA cases from 2012 to 2014 and a characterization of MRSA isolates, including phage-, spa- and PFGE-typing. Additionally, MRSA screening was performed from the hospital staff and the environment. To identify the reason for the outbreak, work-related, psychological and behavioral factors were investigated by impartial audits and staff interviews. RESULTS: Thirty-one MRSA cases were registered during the study period, and 36 isolates were investigated. Molecular typing determined the outbreak strain (phage type 54/812, PFGE type A4, spa type t003) and identified the probable index case. Nasal carriage in one employee and a high environmental contamination with the outbreak strain was documented. Important gaps in nursing procedures and general management were identified. Elevated stress levels and communication problems preceded the outbreak. Compliance with hand hygiene and isolation procedures was evaluated as appropriate. CONCLUSION: This study demonstrates the complexity of controlling hospital-associated infections. The combined use of different typing methods is beneficial for outbreak investigations. Psychological, behavioral and other work-related factors have an important impact on the spread of nosocomial pathogens. These factors should be addressed and integrated in routine infection control practice.

• MeSH
• chování MeSH
• epidemický výskyt choroby * statistika a číselné údaje   MeSH
• hospitalizace statistika a číselné údaje   MeSH
• kontrola infekce metody   statistika a číselné údaje   MeSH
• lidé MeSH
• methicilin rezistentní Staphylococcus aureus izolace a purifikace   fyziologie   MeSH
• práce psychologie   MeSH
• průzkumy a dotazníky MeSH
• stafylokokové infekce epidemiologie   mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Polymerase chain reaction (PCR) amplification of antibiotic resistance genes as well as staphylococcal cassette chromosome mec (SCCmec) typing and pulsed-field gel electrophoresis (PFGE) of SmaI macrorestriction fragments of genomic DNA were used to characterize 45 methicillin-resistant coagulase-negative staphylococci (MRCoNS) isolates responsible of bacteremia recovered in patients at the Bone Marrow Transplant Centre of Tunisia in 1998-2007. Among the 45 MRCoNS isolates, Staphylococcus epidermidis was the most prevalent species (75.6%) followed by Staphylococcus haemolyticus (22.2%) and Staphylococcus hominis (2.2%). Extended susceptibility profiles were generated for MRCoNS against 16 antimicrobial agents. Out of 45 mecA-positive strains, 43 (95.6%) were phenotypically methicillin-resistant and two (4.4%) were methicillin-susceptible. The msr(A) was the most prevalent gene (13 isolates; 48.1%) among erythromycin-resistant isolates. The erm(C) was found alone in seven (25.9%) or in combination with both erm(A) and erm(B) in two (7.4%) isolates. The aac(6')-Ie-aph(2″)-Ia was the most prevalent gene among aminoglycoside-resistant isolates, detected alone in 14 isolates (33.3%) isolates, in combination with ant(4')-Ia in 18 (42.8%) isolates, in combination with aph(3')-IIIa in four (9.5%) or with both ant(4')-Ia and aph(3')-IIIa in two (4.7%) isolates. The ant(4')-Ia was detected in three (7.1%) isolates and the aph(3')-IIIa in one (2.4%) isolate. Among tetracycline-resistant isolates, six (85.7%) strains harbored the tet(K) gene and one (14.3%) strain carried tet(K) and tet(M) genes. SCCmec types IV (31%) and III (24.5%), the most prevalent types detected, were found to be more resistant to non-β-lactam antibiotics. A wide diversity of isolates was observed by PFGE among MRCoNS.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální geny MeSH
• bakteriální léková rezistence MeSH
• bakteriemie mikrobiologie   MeSH
• DNA bakterií genetika   metabolismus   MeSH
• hematologické nádory komplikace   MeSH
• koagulasa metabolismus   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• molekulární typizace MeSH
• polymerázová řetězová reakce metody   MeSH
• pulzní gelová elektroforéza MeSH
• restrikční endonukleasy typu II metabolismus   MeSH
• rezistence na methicilin * MeSH
• stafylokokové infekce mikrobiologie   MeSH
• Staphylococcus epidermidis klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• Staphylococcus haemolyticus klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• Staphylococcus hominis klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• techniky typizace bakterií MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Tunisko MeSH
• Názvy látek
• antibakteriální látky MeSH
• CCCGGG-specific type II deoxyribonucleases MeSH Prohlížeč
• DNA bakterií MeSH
• koagulasa MeSH
• restrikční endonukleasy typu II MeSH

High occurrence of the non-macrolide-lincosamide-streptogramin B resistance genes msrA (53%) and linA/linA' (30%) was found among 98 methicillin-resistant coagulase-negative staphylococci additionally resistant to macrolides and/or lincosamides. The gene msrA predominated in Staphylococcus haemolyticus (43 of 62 isolates). In Staphylococcus epidermidis, it was present in 7 of 27 isolates. A novel mechanism of resistance to lincosamides appears to be present in 10 genetically related isolates of S. haemolyticus in the absence of ermA, ermC, msrA, and linA/linA'.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence * genetika   MeSH
• bakteriální proteiny genetika   MeSH
• incidence MeSH
• koagulasa metabolismus   MeSH
• lidé MeSH
• linkosamidy MeSH
• makrolidy farmakologie   MeSH
• membránové transportní proteiny genetika   MeSH
• mikrobiální testy citlivosti MeSH
• rezistence na methicilin * MeSH
• Staphylococcus klasifikace   účinky léků   enzymologie   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• antibakteriální látky MeSH
• bakteriální proteiny MeSH
• koagulasa MeSH
• linkosamidy MeSH
• makrolidy MeSH
• membránové transportní proteiny MeSH
• msrA protein, Staphylococcus epidermidis MeSH Prohlížeč

Bacterial species of the genus Staphylococcus known as important human and animal pathogens are the cause of a number of severe infectious diseases. Apart from the major pathogen Staphylococcus aureus, other species until recently considered to be nonpathogenic may also be involved in serious infections. Rapid and accurate identification of the disease-causing agent is therefore prerequisite for disease control and epidemiological surveillance. Modern methods for identification and typing of bacterial species are based on genome analysis and have many advantages compared to phenotypic methods. The genotypic methods currently used in molecular diagnostics of staphylococcal species, particularly of S. aureus, are reviewed. Attention is also paid to new molecular methods with the highest discriminatory power. Efforts made to achieve interlaboratory reproducibility of diagnostic methods are presented.

• MeSH
• genotyp MeSH
• lidé MeSH
• polymerázová řetězová reakce MeSH
• rezistence na methicilin MeSH
• sekvenční analýza DNA MeSH
• Staphylococcus klasifikace   genetika   izolace a purifikace   MeSH
• techniky typizace bakterií MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

The purpose of our study was the molecular characterization of methicillin-resistant Staphylococcus aureus (MRSA) strains isolated in 21 hospitals in the Czech Republic in the period 2000-2002 and comparison with previous results from 1996-1997. Strains were analyzed by pulsed-field gel electrophoresis (PFGE) of SmaI digests and ribotyping of HindIII digests hybridized with a 16S-23S DNA probe. The prevalence of the most clinically important macrolide (ermA, ermB, ermC, and msrA) and aminoglycoside (aph3', ant4', and aac6'-aph2") resistance genes was evaluated as well. Selected isolates representative of each clonal type were analyzed by multilocus sequence typing and by a multiplex PCR method capable of identifying the structural type of the staphylococcal cassette chromosome mec (SCCmec) carried by the bacteria. Our results document the displacement of the Brazilian clone (ST239, SCCmec type IIIA, PFGE type B, ribotype H1) by a new clone that we named "Czech clone" (ST239, SCCmec type IIIA, PFGE type F, ribotype H6) and the maintenance of the Iberian clone (ST247, SCCmec type IA, PFGE type A, ribotype H2) exclusively in one hospital in the Czech Republic. In addition, we found a correlation between the distribution of aminoglycoside resistance genes and MRSA clonal types.

• MeSH
• lidé MeSH
• makrolidy farmakologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• pulzní gelová elektroforéza MeSH
• rezistence na methicilin genetika   MeSH
• stafylokokové infekce mikrobiologie   MeSH
• Staphylococcus aureus klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• zeměpis MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• makrolidy MeSH