• MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• klostridiové infekce mikrobiologie   MeSH
• lidé MeSH
• polymerázová řetězová reakce MeSH
• ribotypizace * MeSH
• tandemové repetitivní sekvence MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• dopisy MeSH
• komentáře MeSH
• Geografické názvy
• Česká republika MeSH
• Polsko MeSH

BACKGROUND: Clostridium difficile infection (CDI) is a major cause of antibiotic-associated diarrhoea. Given an increasing CDI incidence and global spread of epidemic ribotypes, a 1-year study was performed to analyse the molecular characteristics of C. difficile isolates and associated clinical outcomes from patients diagnosed with CDI in the Internal Medicine department at University Hospital Motol, Prague from February 2013 to February 2014. RESULTS: A total of 85 unformed stool samples were analysed and CDI was laboratory confirmed in 30 patients (6.8 CDI cases per 10,000 patient bed days and 50.6 CDI cases per 10,000 admissions). The CDI recurrence rate within 3 months of treatment discontinuation was 13.3% (4/30). Mortality within 3 months after first CDI episode was 26.7% (8/30), with CDI the cause of death in two cases. 51.9% of C. difficile isolates belonged to PCR-ribotype 176. MLVA of ribotype 176 isolates revealed two clonal complexes formed by 10/14 isolates. ATLAS scores and Horn's index were higher in patients with ribotype 176 infections than with non-ribotype 176 infections. CONCLUSION: This study highlights the clinical relevance of C. difficile PCR-ribotype 176 and its capacity to spread within a healthcare facility.

• MeSH
• analýza přežití MeSH
• antibakteriální látky aplikace a dávkování   škodlivé účinky   MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• feces mikrobiologie   MeSH
• klostridiové infekce epidemiologie   mikrobiologie   mortalita   patologie   MeSH
• lidé MeSH
• nemocnice univerzitní MeSH
• polymerázová řetězová reakce MeSH
• průjem chemicky indukované   epidemiologie   mikrobiologie   patologie   MeSH
• recidiva MeSH
• ribotypizace * MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Clostridium difficile is a major nosocomial pathogen of present times. The analysis of 624 C. difficile strains from 11 hospitals in the Czech Republic in 2013 revealed that 40% of isolates belonged to ribotype 176. These results suggest that the incidence of CDI (C. difficile infection) in the Czech Republic has increased probably in connection with C. difficile ribotype 176. The molecular systems Xpert C. difficile Epi assay (Cepheid Inc., Sunnyvale, CA) diagnoses toxigenic strains and supports C. difficile ribotype 027 determination based on three specific target places in the toxigenic C. difficile genome. Twenty-nine strains cultivated from stool specimens were evaluated by the Xpert systems as presumed C. difficile PCR ribotype 027 were confirmed as a C. difficile ribotype 176 based on ribotyping. A further 120 C. difficile strains of ribotype 176 were examined for the presence of genes tcdB, cdtB and deletion in position 117 in the tcdC gene. Our experience shows that due to the correspondence of the target places, C. difficile ribotype 176 may be interpreted as ribotype 027 by Xpert C. difficile Epi assay (Cepheid Inc., Sunnyvale, CA). Further molecular analysis as ribotyping based on capillary electrophoresis is needed to differentiate between C. difficile ribotypes 027 and 176 for appropriate epidemiological situation control on local and national levels.

• MeSH
• bakteriální proteiny genetika   MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• delece genu MeSH
• feces mikrobiologie   MeSH
• infekce spojené se zdravotní péčí mikrobiologie   MeSH
• lidé MeSH
• nemocnice statistika a číselné údaje   MeSH
• ribotypizace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Antibiotic profiling of twenty Czech Clostridium difficile PCR-ribotype 176 isolates revealed a high level of resistance to erythromycin, ciprofloxacin and moxifloxacin (n = 20) and to rifampicin (n = 13). Accumulation of resistance mechanisms to multiple antibiotics highlight that PCR-ribotype 176 belong to problematic epidemic strains.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální proteiny genetika   metabolismus   MeSH
• Clostridioides difficile klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• lidé MeSH
• mnohočetná bakteriální léková rezistence * MeSH
• molekulární sekvence - údaje MeSH
• mutace MeSH
• pseudomembranózní enterokolitida mikrobiologie   MeSH
• ribotypizace MeSH
• sekvence nukleotidů MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

The first case of Clostridium difficile RT027 infection in the Czech Republic (CZ) was identified. The patient had been hospitalised in Germany prior to moving to CZ. Multiple-Locus Variable number tandem repeat Analysis revealed a genetic relatedness between the patient's isolate and RT027 isolate collected in the German hospital.

• MeSH
• cestování * MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• hospitalizace MeSH
• klostridiové infekce diagnóza   mikrobiologie   MeSH
• lidé MeSH
• minisatelitní repetice MeSH
• polymerázová řetězová reakce * MeSH
• ribotypizace * MeSH
• senioři nad 80 let MeSH
• Check Tag
• lidé MeSH
• senioři nad 80 let MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH
• Německo MeSH

In 2014, 18 hospitals in the Czech Republic participated in a survey of the incidence of Clostridium difficile infections (CDI) in the country. The mean CDI incidence was 6.1 (standard deviation (SD):7.2) cases per 10,000 patient bed-days and 37.8 cases (SD: 41.4) per 10,000 admissions. The mean CDI testing frequency was 39.5 tests (SD: 25.4) per 10,000 patient bed-days and 255.8 tests (SD: 164.0) per 10,000 admissions. A total of 774 C. difficile isolates were investigated, of which 225 (29%) belonged to PCR ribotype 176, and 184 isolates (24%) belonged to PCR ribotype 001. Multilocus variable-number tandem repeat analysis (MLVA) revealed 27 clonal complexes formed by 84% (190/225) of PCR ribotype 176 isolates, and 14 clonal complexes formed by 77% (141/184) of PCR ribotype 001 isolates. Clonal clusters of PCR ribotypes 176 and 001 were observed in 11 and 7 hospitals, respectively. Our data demonstrate the spread of two C. difficile PCR ribotypes within 18 hospitals in the Czech Republic, stressing the importance of standardising CDI testing protocols and implementing mandatory CDI surveillance in the country.

• MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• elektroforéza kapilární MeSH
• feces mikrobiologie   MeSH
• incidence MeSH
• klostridiové infekce diagnóza   epidemiologie   MeSH
• lidé MeSH
• minisatelitní repetice MeSH
• nemocnice statistika a číselné údaje   MeSH
• oznamovací povinnost MeSH
• polymerázová řetězová reakce MeSH
• průjem epidemiologie   mikrobiologie   MeSH
• ribotypizace metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH

PURPOSE: The objective of this survey was to determine the incidence of Clostridium difficile infections (CDI) at the Department of Infectious Diseases, Bulovka Hospital, and to evaluate clinical and epidemiological data on CDI patients together with a detailed molecular characterisation of C. difficile isolates. The patient outcomes were correlated to causative C. difficile PCR-ribotype. METHODS: The twelve-month study (2013) comprised patients two years of age and older with CDI. CDI severity was estimated using ESCMID criteria and ATLAS scoring. C. difficile isolates were further characterized using ribotyping, Multiple-Locus Variable Tandem-Repeats analysis (MLVA) and investigation of antibiotic-resistance determinants (gyrA, gyrB, rpoB, ermB). RESULTS: A total of 619 diarrhoeal stools were investigated. Seventy-two stool samples were GDH and toxin A/B positive, and 39 samples were GDH positive only and subsequently toxigenic C. difficile was cultured. In total, 111 C. difficile isolates were characterized, of which 64 (57.7%) belonged to PCR-ribotype 176. MLVA analysis of PCR-ribotype 176 isolates revealed 11 clonal complexes. Seventy-two isolates (64.9%) showed amino acid substitution Thr82Ile in the GyrA, and sixty-two isolates (55.9%) showed amino acid substitutions Arg505Lys together with His502Asn, or Asp492Glu together with Arg505Lys in the RpoB. Twelve isolates (10.8%) were ermB positive. Severe CDI according to the ESCMID criteria was recorded in forty-two patients (37.8%), and sixteen patients (14.4%) had ATLAS score ≥ 6. Twenty-nine patients (26.1%) had recurrent CDI and twenty-four patients (21.6%) died during the study period. CONCLUSIONS: A higher rate of severe CDI, recurrences and mortality in association with PCR-ribotype 176 infections were observed. The high incidence of PCR-ribotype 176 in the study, and the presence of clonal relatedness between PCR-ribotype 176 isolates, indicate its higher capacity to spread in a hospital setting, which in turn highlights the need to implement strict epidemic measures when PCR-ribotype 176 occurs.

• MeSH
• analýza přežití MeSH
• antibakteriální látky terapeutické užití   MeSH
• bakteriální léková rezistence genetika   MeSH
• bakteriální proteiny genetika   MeSH
• Clostridioides difficile klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• dítě MeSH
• dospělí MeSH
• exprese genu MeSH
• infekce spojené se zdravotní péčí diagnóza   farmakoterapie   mortalita   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• multilokusová sekvenční typizace MeSH
• mutace MeSH
• předškolní dítě MeSH
• průjem diagnóza   farmakoterapie   mortalita   patologie   MeSH
• pseudomembranózní enterokolitida diagnóza   farmakoterapie   mortalita   patologie   MeSH
• retrospektivní studie MeSH
• ribotypizace MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• stupeň závažnosti nemoci MeSH
• výsledek terapie MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Cíl práce: Porovnání senzitivity a specificity dvou komerčních testů na stanovení Clostridium difficile GDH (glutamát dehydrogenázy) a toxinů A/B. Materiál a metodiky: Bylo testováno 86 stolic pacientů hospitalizovaných ve FN Motol. GDH a toxiny A/B byly paralelně vyšetřovány dvěma testy: C. difficile Quik Chek Complete?, Techlab,USA) a Liaison? C. difficile GDH a Toxins AαB (DiaSorin USA). Současně byly ze stolic izolovány nukleové kyseliny pomocí UltraClean? Fecal DNA (MoBio Laboratories, USA). Pro metodu PCR byla použita komerční souprava C. difficile Elite MGB? kit (Nanogen, Itálie). Anaerobní kultivace na selektivním médiu pro C. difficile (Oxoid) proběhla u všech vzorků vykazující pozitivitu alespoň jednoho testu. Čisté izoláty byly molekulárně analyzovány ribotypizací. Výsledky: GDH a toxin A/B negativních oběma metodami bylo 36 vzorků (42 %). GDH a toxin pozitivních oběma metodami bylo 20 vzorků (23 %). GDH pozitivních a toxin negativních oběma metodami, ale PCR pozitivních bylo 9 vzorků (10 %). GDH pozitivních a toxin negativních oběma metodami a PCR negativních bylo 11 vzorků (13 %). GDH pozitivní, toxin pozitivní pouze testem Liaison? bylo 6 vzorků (7 %). GDH pozitivní pouze Liaison? byly 4 vzorky (5 %). K selhání kultivace došlo u 11 vzorků (13), z toho u 7 vzorků byl pozitivní test PCR. PCR reakce byla inhibována v 5 případech (6 %). Zachycené ribotypy toxigenní: AI-3, 001, 002, 012,014, 017, 020, 049, 054, 078, 176, 203, 413 a netoxigenní: AI-34, AI-61, 010, 485, 495, 596. Závěr: Stanovení toxinu A/B testem Liaison? vykazuje u námi vyšetřeného souboru o 7 % vyšší senzitivitu. Dvoukrokové uspořádání testů také přináší ekonomickou úsporu, která může být využita např. k zavedení PCR metod do diagnostického algoritmu laboratoře.

Study objective: Comparison of two commercially avail­able tests for the detection of Clostridium difficile Glutamate Dehydrogenase (GDH) and toxins A and B for their sensitivity and specificity. Material and methods: Eighty-six stool samples from patients hospitalised in the Motol University Hospital were analysed. GDH and toxins A and B were assayed in parallel by two tests: C. difficile Quik Chek Complete? (Techlab, USA) and Liaison? C. difficile GDH and Toxins AαB (DiaSorin, USA). From the stool samples, nucleic acids were also isolated using the UltraClean? Fecal DNA kit (MoBio Laboratories, USA). The commercially available C. difficile Elite MGB? kit (Nanogen, Italy) was used for the polymerase chain reaction (PCR). Anaerobic culture on C. difficile selective medium (Oxoid) was performed for all positive samples at least in one test. Pure isolates were characterized by PCR ribotyping. Results: Thirty-six (42%) samples were GDH negative and toxin A/B negative by both tests. Twenty (23%) samples were GDH positive and toxin A/B positive by both tests. Nine (10%) samples were GDH positive and toxin negative by both tests, but were positive by PCR. Eleven (13%) samples that were GDH positive and toxin negative by both tests remained negative by PCR. Six (7%) samples only were GDH positive and toxin positive by the Liaison? test alone. Four (5%) samples were GDH-positive by theLiaison? test alone. Culture failure was observed in 11 (13%) samples, of which seven were positive by PCR. PCR was inhibited in five (6%) samples. The following toxigenic ribotypes: AI-3, 001, 002, 012,014, 017, 020, 049, 054, 078, 176, 203, and 413 and non-toxigenic ribotypes: AI-34, AI-61, 010, 485, 495, and 596 were identified. Conclusion: The Liaison? test had seven percent higher sensitivity for the detection of toxins A/B. The two-step protocol of the tests is also cost-saving. The savings can be used e.g. for incorporating the PCR techniques into the diagnostic algorithm of the laboratory.

• MeSH
• antigeny bakteriální analýza   MeSH
• Clostridioides difficile * enzymologie   chemie   izolace a purifikace   MeSH
• feces mikrobiologie   MeSH
• glutamátdehydrogenasa imunologie   MeSH
• imunoenzymatické techniky metody   využití   MeSH
• klostridiové infekce * diagnóza   genetika   mikrobiologie   MeSH
• lidé MeSH
• polymerázová řetězová reakce MeSH
• senzitivita a specificita MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH

This study aimed to implement a toxigenic culture as an optional third diagnostic step for glutamate dehydrogenase (GDH)-positive and toxin A/B-negative diarrheal stool samples into a diagnostic algorithm for Clostridioides (Clostridium) difficile infection (CDI), and to characterise C. difficile isolates for epidemiological purposes. During the 5-month study, 481 diarrhoeal stool samples from three Slovak hospitals were investigated and 66 non-duplicated GDH-positive stool samples were found. Of them, 36 were also toxin A/B-positive. Twenty-three GDH-positive and toxin A/B-negative stool samples were shown subsequently to be positive following toxigenic culture (TC). Molecular characterisation of C. difficile isolates showed the predominance of PCR ribotype (RT) 001 (n = 37, 56.1%) and the occurrence of RT 176 (n = 3, 4.5%). C. difficile RT 001 isolates clustered to eight clonal complexes (CCs) using multiple-locus variable-number tandem repeats analysis (MLVA). Interestingly, one third of RT 001 isolates clustering in these CCs were cultured from toxin A/B-negative stool samples. Our observations highlight the need of use multiple step diagnostic algorithm in CDI diagnosis in order to detect all CDI cases and to avoid the spread of C. difficile in healthcare settings.

• MeSH
• algoritmy MeSH
• bakteriální proteiny analýza   MeSH
• Clostridioides difficile klasifikace   izolace a purifikace   MeSH
• diagnostické techniky molekulární MeSH
• enterotoxiny analýza   nedostatek   MeSH
• feces mikrobiologie   MeSH
• glutamátdehydrogenasa analýza   MeSH
• klostridiové infekce diagnóza   epidemiologie   mikrobiologie   MeSH
• lidé MeSH
• minisatelitní repetice genetika   MeSH
• molekulární typizace * MeSH
• multilokusová sekvenční typizace MeSH
• nemocnice MeSH
• polymerázová řetězová reakce * MeSH
• průjem diagnóza   epidemiologie   mikrobiologie   MeSH
• ribotypizace metody   MeSH
• shluková analýza MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• Geografické názvy
• Slovenská republika MeSH

PURPOSE: To characterise and compare twenty-eight Finnish Clostridium difficile RT027-like isolates, selected based on the presence of 18 bp deletion in the tcdC gene and toxin gene profile (A, B, binary), with eleven RT027 isolates from different Finnish geographical areas and time periods. METHODS: Twenty-eight C. difficile RT027-like isolates and 11 RT027 comparative strains were characterised by capillary-electrophoresis (CE) ribotyping, multi-locus variable tandem-repeats analysis (MLVA), multi-locus sequence typing (MLST), and sequencing of tcdC and gyrA gene fragments. Susceptibility to moxifloxacin was determined by E-test. RESULTS: Of 28 RT027-like isolates, seven RTs (016, 034, 075, 080, 153, 176 and 328), three WEBRIBO types (411, 475, AI-78) and three new profiles (F1-F3) were identified. MLVA revealed six clonal complexes (RTs 016, 027, 176 and F3). MLST showed eleven sequence types (1, 41, 47, 67, 95, 191,192, 223, 229, 264 and new ST). Twenty-two isolates (RTs 016, 080, 176, 328, F1, F2, F3 and WRTAI-78) carried Δ117 in the tcdC gene. Isolates of RTs 016, 027 and 176 were moxifloxacin resistant and harboured Thr82Ile in the GyrA. CONCLUSION: Our results show a high diversity within 28 Finnish RT027-like C. difficile isolates, with twelve CE-ribotyping profiles and eleven STs. MLVA revealed the regional spread of RTs 016, 027, 176 and F3. The presence of Δ117 in the tcdC gene in eight non-027 RTs highlights the importance of careful interpretation of the results from molecular systems targeting this site in the genome of C. difficile and the need of strain typing for epidemiological purposes.

• MeSH
• ADP-ribosatransferasy genetika   MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence MeSH
• bakteriální proteiny genetika   MeSH
• bakteriální toxiny analýza   genetika   MeSH
• Clostridioides difficile klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• DNA bakterií analýza   MeSH
• DNA gyráza genetika   MeSH
• dospělí MeSH
• enterotoxiny genetika   MeSH
• fluorochinolony farmakologie   MeSH
• genotyp MeSH
• klostridiové infekce epidemiologie   mikrobiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• minisatelitní repetice genetika   MeSH
• mladý dospělý MeSH
• molekulární epidemiologie MeSH
• moxifloxacin MeSH
• multilokusová sekvenční typizace metody   MeSH
• polymerázová řetězová reakce metody   MeSH
• represorové proteiny genetika   MeSH
• ribotypizace metody   MeSH
• sekvence nukleotidů MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• techniky typizace bakterií * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Finsko MeSH