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

AIM: To perform a retrospective analysis of the high occurrence of Clostridium difficile infection in the surgical department of a Czech tertiary care hospital and to identify weaknesses in C. difficile infection (CDI) prevention and control policies. METHODS: Clinical and epidemiological data on eleven CDI cases were collected. C. difficile isolates were characterized by capillary electrophoresis ribotyping, multilocus variable tandem repeat analysis (MLVA), gyrA gene fragment sequencing, and erm(B) fragment PCR amplification. Antibiotic susceptibility to metronidazole, vancomycin, ciprofloxacin, moxifloxacin, and clindamycin was tested. FINDINGS: Eleven CDI cases were caused by C. difficile PCR ribotype 001 strains. These strains revealed two different MLVA profiles with 11 tandem repeat differences. All isolates were susceptible to metronidazole and vancomycin and resistant to ciprofloxacin (MIC ≥32 mg/L), moxifloxacin (MIC ≥32 mg/L), and clindamycin (MIC ≥256 mg/L). All isolates revealed amino acid substitution Thr82Ile, in the GyrA and were erm(B) negative. CONCLUSION: Two fluoroquinolone and clindamycin-resistant C. difficile PCR ribotype 001 strain clusters occurred at one of the surgical departments of a tertiary care hospital. Ineffective decontamination with suboptimal concentration and time of exposure of sporicidal disinfectants may have resulted in C. difficile transmission.

• MeSH
• antibakteriální látky farmakologie   MeSH
• centra terciární péče MeSH
• ciprofloxacin farmakologie   MeSH
• Clostridioides difficile klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• DNA bakterií genetika   metabolismus   MeSH
• DNA gyráza genetika   metabolismus   MeSH
• elektroforéza kapilární MeSH
• exprese genu MeSH
• fluorochinolony farmakologie   MeSH
• klindamycin farmakologie   MeSH
• klostridiové infekce farmakoterapie   mikrobiologie   MeSH
• lidé MeSH
• methyltransferasy genetika   metabolismus   MeSH
• metronidazol farmakologie   MeSH
• mikrobiální testy citlivosti MeSH
• minisatelitní repetice MeSH
• mnohočetná bakteriální léková rezistence genetika   MeSH
• multilokusová sekvenční typizace MeSH
• retrospektivní studie MeSH
• ribotypizace MeSH
• senioři MeSH
• substituce aminokyselin MeSH
• vankomycin farmakologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH

PURPOSE: The purpose of this study was to determine the incidence of Clostridium difficile infections (CDI) and to characterise the isolates in 14 departments of ten academic hospitals in Slovakia. METHODS: During a one-month study (September 2012) all unformed stool samples were investigated using a rapid test to detect the presence of GDH and toxins A/B. Positive samples were cultured anaerobically and C. difficile isolates were characterised by ribotyping, multiple-locus variable-number tandem repeats analysis, and gyrA, rpoB and ermB investigation. RESULTS: A total of 194 unformed stool samples were investigated and 38 (19.6 %) had a positive rapid test. Of 38 samples, 27 revealed a positive result for GDH and free toxins A/B in the stool, and 11 samples only for the presence of GDH. The mean CDI incidence in 2012 was 5.2 cases per 10,000 patient bed-days. Twenty C. difficile isolates were available for molecular analysis; seventeen belonged to PCR-ribotype 001 (85 %) whereas the remaining three isolates were identified as PCR-ribotypes 017, 078 and 449. MLVA of the PCR-ribotype 001 isolates identified two clonal complexes and a close genetic relatedness between isolates from six different hospitals. Molecular analysis of antibiotic-resistance determinants revealed the presence of ermB gene encoding resistance to the MLSB group of antibiotics in 90 % of isolates, and Thr82Ile amino acid substitution in the gyrA gene associated with resistance to fluoroquinolones in 85 % of isolates. CONCLUSIONS: We conclude that C. difficile PCR-ribotype 001 is the predominant PCR-ribotype in Slovakia with a strong potential for clonal spread and development of multidrug resistance.

• MeSH
• bakteriální proteiny genetika   MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• dospělí MeSH
• incidence MeSH
• klostridiové infekce epidemiologie   mikrobiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• minisatelitní repetice MeSH
• mladý dospělý MeSH
• molekulární epidemiologie MeSH
• nemocnice univerzitní MeSH
• polymerázová řetězová reakce MeSH
• průjem chemicky indukované   epidemiologie   mikrobiologie   MeSH
• retrospektivní studie MeSH
• ribotypizace * MeSH
• senioři nad 80 let MeSH
• senioři 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
• práce podpořená grantem MeSH
• Geografické názvy
• Slovenská republika MeSH

Extra-intestinal infections caused by Clostridium difficile are rare. The risk of extra-intestinal infections associated with C. difficile may be particularly relevant in environments contaminated with C. difficile spores. This paper describes the case of a non-diarrheic patient colonized with C. difficile ribotype 014 in the intestinal tract who developed a post-surgical wound infection by C. difficile ribotype 078. The infection responded to metronidazole administered first intravenously and then orally. This case indicates that C. difficile may not only be related to diarrheic diseases, but also to infections of non-healing wounds, especially in situations when C. difficile is the only isolated pathogen.

• MeSH
• antibakteriální látky aplikace a dávkování   MeSH
• Clostridioides difficile klasifikace   genetika   růst a vývoj   izolace a purifikace   MeSH
• dospělí MeSH
• infekce chirurgické rány farmakoterapie   mikrobiologie   MeSH
• klostridiové infekce farmakoterapie   mikrobiologie   MeSH
• lidé MeSH
• metronidazol aplikace a dávkování   MeSH
• ribotypizace MeSH
• střeva mikrobiologie   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

We aimed to characterize Clostridioides difficile isolates cultured during a six-month single-center study from stool samples of patients with C. difficile infection (CDI) genotyped by the Xpert®C. difficile/Epi assay by polymerase chain reaction (PCR) ribotyping, toxin genes' detection and multi-locus variable number tandem repeats analysis (MLVA). The susceptibility to metronidazole, vancomycin and moxifloxacin was determined by agar dilution. In addition, the presence of Thr82Ile in the GyrA and a single nucleotide deletion at position (Δ117) in the tcdC gene were investigated. Between January 1 and June 30, 2016, of 114 CDIs, 75 cases were genotyped as presumptive PCR ribotype (RT) 027 infections using a commercial assay. C. difficile isolates cultured from presumptive RT027 stool samples belonged to RT176. These isolates carried genes for toxin A (tcdA), B (tcdB), binary (cdtA/B) and had Δ117 in the tcdC gene. Using MLVA, the 71/75 isolates clustered into two clonal complexes (CCs). Of these, 39 isolates (54.9%) were from patients hospitalized in acute care and 32 isolates (45.1%) were isolated from patients hospitalized in the long-term care department. All isolates were susceptible to metronidazole and vancomycin, and 105 isolates were resistant to moxifloxacin (92%) carrying Thr83Ile in the GyrA. An outbreak of RT176 CDIs, suspected as RT027, was recognized in a Slovakian hospital. In order to monitor the emergence and spread of RT027-variants, the identification of a presumptive RT027 CDI should be confirmed at a strain level by PCR ribotyping.

• Publikační typ
• časopisecké články MeSH

• 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

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

Clostridium difficile je závažným nozokomiálním patogenem současnosti uváděným hlavně v souvislosti s postantibiotickými průjmy. Bližší určení klinicky významných izolátů C. difficile přispívá ke zmapování epidemiologické situace v ČR. Celkem 624 izolátů C. difficile z 11 mikrobiologických pracovišť bylo typizováno pomocí ribotypizace založené na kapilární elektroforéze. Celkem bylo identifikováno 75 různých ribotypů. 40% izolátů příslušelo k ribotypu 176. Dalšími častěji se opakujícími ribotypy byly: 001, 012, 014, 017, 020 a 078.

Clostridium difficile is a major nosocomial pathogen reported mainly in the context of post-antibiotic diarrhoea. Further identification of clinically significant isolates of C. difficile contributes to the mapping of the epidemiological situation in the Czech Republic. A total of 624 C. difficile isolates from 11 microbiological centres were characterized using capillary gel electrophoresis-based PCR ribotyping. Forty percent of the isolates belonged to ribotype 176. Other frequently identified ribotypes were: 001, 012, 014, 017, 020, and 078.

• Klíčová slova
• surveillance,
• MeSH
• Clostridioides difficile * izolace a purifikace   patogenita   růst a vývoj   MeSH
• elektroforéza kapilární MeSH
• lidé MeSH
• průjem prevence a kontrola   MeSH
• ribotypizace metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika 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