The Pseudomonas aeruginosa population has a nonclonal epidemic structure. It is generally composed of a limited number of widespread clones selected from a background of many rare and unrelated genotypes recombining at high frequency. Due to the increasing prevalence of nosocomial infections caused by multidrug-resistant/extensively drug-resistant (MDR/XDR) strains, it is advisable to implement infection control measures. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) are considered the gold standard methods in bacterial typing, despite being limited by cost, staff, and instrumental demands. Here, we present a novel mini-MLST scheme for P. aeruginosa rapid genotyping based on high-resolution melting analysis. Using the proposed mini-MLST scheme, 3,955 existing sequence types (STs) were converted into 701 melting types (MelTs), resulting in a discriminatory power of D = 0.993 (95% confidence interval [CI], 0.992 to 0.994). Whole-genome sequencing of 18 clinical isolates was performed to support the newly designed mini-MLST scheme. The clonal analysis of STs belonging to MelTs associated with international high-risk clones (HRCs) performed by goeBURST software revealed that a high proportion of the included STs are highly related to HRCs and have also been witnessed as responsible for serious infections. Therefore, mini-MLST provides a clear warning for the potential spread of P. aeruginosa clones recognized as MDR/XDR strains with possible serious outcomes. IMPORTANCE In this study, we designed a novel mini-MLST typing scheme for Pseudomonas aeruginosa. Its great discriminatory power, together with ease of performance and short processing time, makes this approach attractive for prospective typing of large isolate sets. Integrating the novel P. aeruginosa molecular typing scheme enables the development and spread of MDR/XDR high-risk clones to be investigated.

• MeSH
• buněčné klony MeSH
• genotyp MeSH
• lidé MeSH
• molekulární epidemiologie metody   MeSH
• multilokusová sekvenční typizace MeSH
• prospektivní studie MeSH
• pseudomonádové infekce * epidemiologie   mikrobiologie   MeSH
• Pseudomonas aeruginosa * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

In 2003 to 2004, the first five VIM-2 metallo-β-lactamase (MBL)-producing Pseudomonas aeruginosa (MPPA) isolates with an In4-like integron, In461 (aadB-blaVIM-2-aadA6), on conjugative plasmids were identified in three hospitals in Poland. In 2005 to 2015, MPPA expanded much in the country, and as many as 80 isolates in a collection of 454 MPPA (∼18%) had In461, one of the two most common MBL-encoding integrons. The organisms occurred in 49 hospitals in 33 cities of 11/16 main administrative regions. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) classified them into 55 pulsotypes and 35 sequence types (STs), respectively, revealing their remarkable genetic diversity overall, with only a few small clonal clusters. S1 nuclease/hybridization assays and mating of 63 representative isolates showed that ∼85% of these had large In461-carrying plasmids, ∼350 to 550 kb, usually self-transmitting with high efficiency (∼10-1 to 10-2 per donor cell). The plasmids from 19 isolates were sequenced and subjected to structural and single-nucleotide-polymorphism (SNP)-based phylogenetic analysis. These formed a subgroup within a family of IncP-2-type megaplasmids, observed worldwide in pseudomonads from various environments and conferring resistance/tolerance to multiple stress factors, including antibiotics. Their microdiversity in Poland arose mainly from acquisition of different accessory fragments, as well as new resistance genes and multiplication of these. Short-read sequence and/or PCR mapping confirmed the In461-carrying plasmids in the remaining isolates to be the IncP-2 types. The study demonstrated a large-scale epidemic spread of multidrug resistance plasmids in P. aeruginosa populations, creating an epidemiological threat. It contributes to the knowledge on IncP-2 types, which are interesting research objects in resistance epidemiology, environmental microbiology, and biotechnology.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny MeSH
• beta-laktamasy genetika   metabolismus   MeSH
• epidemie * MeSH
• fylogeneze MeSH
• infekce spojené se zdravotní péčí * epidemiologie   MeSH
• integrony genetika   MeSH
• lidé MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice MeSH
• pseudomonádové infekce * farmakoterapie   epidemiologie   MeSH
• Pseudomonas aeruginosa genetika   metabolismus   MeSH
• pulzní gelová elektroforéza MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Polsko MeSH

INTRODUCTION: Toxic epidermal necrolysis (TEN) is a rare, life-threatening autoimmune disease predominantly manifested in the skin and mucous membranes. Today, infectious complications have the dominant share in mortality of TEN patients. Due to the nature of the therapy and administration of immunosuppressive medications, a wide range of potentially pathogenic microorganisms, which cause infectious complications in different compartments in these patients, is not surprising. MATERIAL AND METHODOLOGY: This is a multicentric study, which included all patients with TEN hospitalized between 2000-2015 in specialized centres in the Czech Republic and Slovakia. The total catchment area was over 12.5 million inhabitants. The actual implementation of the project was carried out using data obtained from the registry CELESTE (Central European LyEll Syndrome: Therapeutic Evaluation), when specific parameters relating to epidemiological indicators and infectious complications in patients with TEN were evaluated in the form of a retrospective analysis. RESULTS: In total, 39 patients with TEN were included in the study (12 patients died, mortality was 31%), who were hospitalized in the monitored period. The median age of patients in the group was 63 years (the range was 4-83 years, the mean was 51 years), the median of the exfoliated area was 70% TBSA (total body surface area) (range 30-100%, mean 67%). SCORTEN was calculated for 38 patients on the day of admission. Its median in all patients was 3 (range 1-6; mean 3). Any kind of infectious complication in the study group was recorded in 33 patients in total (85%). In total, 30 patients (77%) were infected with gram-positive cocci, 27 patients (69%) with gram-negative rods, and yeast cells or fibrous sponge were cultivated in 12 patients (31%). A total of 32 patients (82%) were found to have infectious complications in the exfoliated area, 15 patients (39%) had lower respiratory tract infections, 18 patients (46%) urinary tract infections and 15 patients (39%) an infection in the bloodstream. The most common potentially pathogenic microorganism isolated in our study group was coagulase neg. Staphylococcus, which caused infectious complications in 24 patients. Enterococcus faecalis/faecium (19 patients), Pseudomonas aeruginosa (17 patients), Staphylococcus aureus (11 patients) and Escherichia coli (11 patients) were other most frequently isolated micro-organisms. CONCLUSION: The published data were obtained from the unique registry of TEN patients in Central Europe. In the first part, we have succeeded in defining the basic epidemiological indicators in the group of patients anonymously included in this registry. The study clearly confirms that infectious complications currently play an essential role in TEN patients, often limiting the chances of survival. The study also shows a high prevalence of these complications in the period after 15days from the start of hospitalization, when most patients already have completely regenerated skin cover.

• MeSH
• aspergilóza epidemiologie   mortalita   MeSH
• bakteriální infekce epidemiologie   mikrobiologie   mortalita   MeSH
• bakteriemie epidemiologie   mikrobiologie   mortalita   MeSH
• dítě MeSH
• dospělí MeSH
• Enterococcus faecalis MeSH
• Enterococcus faecium MeSH
• infekce močového ústrojí epidemiologie   mikrobiologie   mortalita   MeSH
• infekce vyvolané Escherichia coli epidemiologie   mortalita   MeSH
• kandidóza epidemiologie   mortalita   MeSH
• katétrové infekce epidemiologie   mikrobiologie   mortalita   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mykózy epidemiologie   mikrobiologie   mortalita   MeSH
• pneumonie epidemiologie   mikrobiologie   mortalita   MeSH
• povrch těla MeSH
• předškolní dítě MeSH
• prevalence MeSH
• proporcionální rizikové modely MeSH
• pseudomonádové infekce epidemiologie   mikrobiologie   mortalita   MeSH
• Pseudomonas aeruginosa MeSH
• registrace * MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• stafylokokové infekce epidemiologie   mikrobiologie   mortalita   MeSH
• Staphylococcus aureus MeSH
• Stevensův-Johnsonův syndrom epidemiologie   mikrobiologie   mortalita   MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Slovenská republika MeSH

The objective of this study was to perform molecular surveillance for assessing the spread of carbapenemase-producing Pseudomonas aeruginosa in Czech hospitals. One hundred thirty-six carbapenemase-producing isolates were recovered from 22 hospitals located throughout the country. Sequence type 357 (ST357) dominated (n = 120) among carbapenemase producers. One hundred seventeen isolates produced IMP-type (IMP-7 [n = 116] and IMP-1 [n = 1]) metallo-β-lactamases (MβLs), 15 produced the VIM-2 MβL, and the remaining isolates expressed the GES-5 enzyme. The blaIMP-like genes were located in three main integron types, with In-p110-like being the most prevalent (n = 115). The two other IMP-encoding integrons (In1392 and In1393) have not been described previously. blaVIM-2-carrying integrons included In59-like, In56, and a novel element (In1391). blaGES-5 was carried by In717. Sequencing data showed that In-p110-like was associated with a Tn4380-like transposon inserted in genomic island LESGI-3 in the P. aeruginosa chromosome. The other integrons were also integrated into the P. aeruginosa chromosome. These findings indicated the clonal spread of ST357 P. aeruginosa, carrying the IMP-7-encoding integron In-p110, in Czech hospitals. Additionally, the sporadic emergence of P. aeruginosa producing different carbapenemase types, associated with divergent or novel integrons, punctuated the ongoing evolution of these bacteria.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální chromozomy chemie   MeSH
• beta-laktamasy genetika   metabolismus   MeSH
• epidemiologické monitorování MeSH
• exprese genu MeSH
• genomové ostrovy MeSH
• genotyp MeSH
• incidence MeSH
• integrony MeSH
• izoenzymy genetika   metabolismus   MeSH
• karbapenemy farmakologie   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• mnohočetná bakteriální léková rezistence genetika   MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice MeSH
• pseudomonádové infekce epidemiologie   mikrobiologie   MeSH
• Pseudomonas aeruginosa účinky léků   enzymologie   genetika   izolace a purifikace   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

BACKGROUND: Economic data pertaining to cystic fibrosis (CF), is limited in Europe generally, and completely lacking in Central and Eastern Europe. We performed an analysis of all direct costs associated with CF relative to key disease features and laboratory examinations. METHODS: A retrospective prevalence-based cost-of-illness (COI) study was performed in a representative cohort of 242 CF patients in the Czech Republic, which represents about 65 % of all Czech CF patients. Medical records and invoices to health insurance companies for reference year 2010 were analyzed. RESULTS: The mean total health care costs were €14,486 per patient, with the majority of the costs going towards medicinal products and devices (€10,321). Medical procedures (€2676) and inpatient care (€1829) represented a much smaller percentage of costs. A generalized linear model showed that the strongest cost drivers, for all cost categories, were associated with patient age and lung disease severity (assessed using the FEV1 spirometric parameter), when compounded by chronic Pseudomonas aeruginosa airway infections. Specifically, maximum total costs are around the age 16 years; a FEV1 increase of 1 % point represented a cost decrease of: 0.9 % (medicinal products), 1.7 % (total costs), 2.8 % (procedures) and 7.0 % (inpatient care). CONCLUSIONS: COI analysis and regression modeling using the most recent data available can provide a better understanding of the overall economic CF burden. A comparison of our results with other methodologically similar studies demonstrates that although overall costs may differ, FEV1 can nonetheless be utilized as a generally transferrable indicator of the relative economic impact of CF.

• MeSH
• cystická fibróza ekonomika   epidemiologie   patofyziologie   MeSH
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• náklady na zdravotní péči statistika a číselné údaje   MeSH
• osobní újma zaviněná nemocí * MeSH
• předškolní dítě MeSH
• prevalence MeSH
• pseudomonádové infekce ekonomika   epidemiologie   MeSH
• regresní analýza MeSH
• retrospektivní studie MeSH
• spirometrie MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

BACKGROUND: Pseudomonas aeruginosa is a major cystic fibrosis (CF) pathogen causing chronic respiratory infections and posing a risk for cross-infection between patients with CF. AIM: To propose an algorithm for long-term surveillance of P. aeruginosa and assess its suitability for monitoring the epidemiological situation at a CF centre with approximately 300 patients. METHODS: Over a nine-year period, over 300 P. aeruginosa isolates from 131 infected patients were tested by multi-locus sequence typing (MLST) and/or random amplified polymorphic DNA (RAPD) assay. FINDINGS: MLST analysis led to the identification of 97 different sequence types which were distributed among 17 RAPD-generated (pseudo)clusters. This indicates that the easy-to-perform RAPD assay is only suitable for intra-individual, not interindividual, strain analyses. No epidemic strains were observed. Longitudinal analysis revealed that 110 of the 131 patients were infected with the same strain over the observation period, whereas 21 patients had a strain replacement or a new infection. Chronic infection was found in 99 of the 131 patients, and the remaining 32 patients met the criteria for intermittent infection (as defined by the Leeds criteria). Eighteen of the 32 patients (56%) with intermittent infection were infected with the same strain for up to nine years. CONCLUSION: The strain type only changed in 16% of 131 patients with chronic or intermittent infection. As many as 56% of patients considered to have intermittent infection were actually chronically infected with the same strain for many years.

• MeSH
• cystická fibróza komplikace   MeSH
• dítě MeSH
• dospělí MeSH
• epidemiologické monitorování * MeSH
• genotyp MeSH
• infekce spojené se zdravotní péčí epidemiologie   přenos   MeSH
• lidé středního věku MeSH
• lidé MeSH
• longitudinální studie MeSH
• mladý dospělý MeSH
• molekulární epidemiologie MeSH
• multilokusová sekvenční typizace MeSH
• předškolní dítě MeSH
• pseudomonádové infekce epidemiologie   přenos   MeSH
• Pseudomonas aeruginosa klasifikace   genetika   izolace a purifikace   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• technika náhodné amplifikace polymorfní DNA MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• předškolní dítě MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• antibakteriální látky farmakologie   MeSH
• beta-laktamasy biosyntéza   genetika   MeSH
• beta-laktamová rezistence genetika   MeSH
• infekce spojené se zdravotní péčí epidemiologie   mikrobiologie   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice fakultní MeSH
• polymerázová řetězová reakce metody   MeSH
• pseudomonádové infekce epidemiologie   mikrobiologie   MeSH
• Pseudomonas genetika   klasifikace   účinky léků   MeSH
• Check Tag
• lidé MeSH
• Geografické názvy
• Česká republika MeSH

• MeSH
• dítě MeSH
• dospělí MeSH
• folikulitida mikrobiologie   MeSH
• hlášení nemocí statistika a číselné údaje   MeSH
• kontrola infekčních nemocí statistika a číselné údaje   MeSH
• lidé MeSH
• pseudomonádové infekce diagnóza   epidemiologie   MeSH
• zdroje nemoci mikrobiologie   MeSH
• znečištění vody MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH

To determine whether the high proportion of antimicrobial resistance among hospital isolates of Pseudomonas aeruginosa in the Czech Republic is associated with the spread of multidrug-resistant clones, we investigated 108 bloodstream isolates collected prospectively in 2007. The isolates originated from 48 hospitals in 36 cities and were serotyped, tested for susceptibility to 10 anti-Pseudomonas agents and studied by multilocus sequence typing, macrorestriction analysis and class 1 integron typing. Forty-five isolates were fully susceptible, while 14 and 49 isolates were resistant to 1-2 and 3-9 agents, respectively. A total of 42 multilocus sequence types (ST) were identified, of which ST235 (serotype O11), ST175 (O4) and ST132 (O6) included 19, 16 and 5 isolates, respectively. These three STs encompassed 40 (82%) of 49 isolates resistant to more than two agents and originated from 29 hospitals in 22 cities. Isolates of the same ST had highly similar macrorestriction patterns. Twelve ST235 isolates harbored an integron variable region with the gene cassette array of aacA7-aadA6-orfD, while 15 ST175 isolates shared a region with the aadB-aadA13 array and all ST132 isolates carried a region with aacA4. A carbapenemase-encoding gene (bla(IMP-7)) was detected in a single strain (ST357). In conclusion, the multidrug resistance of Czech P. aeruginosa bloodstream isolates in 2007 was predominantly associated with three epidemic clones, one of which belongs to international clonal complex CC235.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální geny MeSH
• bakteriemie epidemiologie   mikrobiologie   MeSH
• DNA bakterií chemie   genetika   MeSH
• DNA fingerprinting MeSH
• genotyp MeSH
• infekce spojené se zdravotní péčí epidemiologie   mikrobiologie   MeSH
• integrony MeSH
• krev mikrobiologie   MeSH
• lidé MeSH
• mnohočetná bakteriální léková rezistence MeSH
• molekulární sekvence - údaje MeSH
• nemocnice MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• pseudomonádové infekce epidemiologie   mikrobiologie   MeSH
• Pseudomonas aeruginosa klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• sekvenční analýza DNA MeSH
• sérotypizace MeSH
• shluková analýza MeSH
• techniky typizace bakterií MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

The aim of the study was to determine the prevalence of Pseudomonas aeruginosa and Klebsiella pneumoniae strains in patients with acute leukemias, to assess their clinical significance, and to define the sources and ways of their spread using genetic analysis. Thirty-four patients were investigated during the observed period. Twenty-one strains of Pseudomonas aeruginosa and 35 strains of Klebsiella pneumoniae were isolated from patient samples. In the case of Pseudomonas aeruginosa, 47.6% of strains were identified as pathogens and caused infection. By contrast, only 4 isolates (11.4%) of Klebsiella pneumoniae could be regarded as etiological agents of bacterial infection. Based on the obtained results, Klebsiella pneumoniae strains are assumed to be of mostly endogenous origin. In the case of Pseudomonas aeruginosa strains, the proportion of identical strains detected in various patients was higher and exogenous sources were more significant. In addition, our results confirmed the ability of Pseudomonas aeruginosa strains to survive on a particular site in the hospital for a longer time.

• MeSH
• antibakteriální látky farmakologie   MeSH
• financování organizované MeSH
• hematologické nádory komplikace   MeSH
• infekce bakteriemi rodu Klebsiella epidemiologie   mikrobiologie   přenos   MeSH
• Klebsiella pneumoniae genetika   izolace a purifikace   klasifikace   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• prevalence MeSH
• pseudomonádové infekce epidemiologie   mikrobiologie   přenos   MeSH
• Pseudomonas aeruginosa genetika   izolace a purifikace   klasifikace   MeSH
• Check Tag
• lidé MeSH