The ribotyping of Clostridioides difficile is one of the basic methods of molecular epidemiology for monitoring the spread of C. difficile infections. In the Czech Republic, this procedure is mainly available in university hospitals. The introduction of ribotyping in a tertiary health care facility such as Liberec Regional Hospital not only increases safety in the facility but also supports regional professional development. In our study, 556 stool samples collected between June 2017 and June 2018 were used for C. difficile infection screening, followed by cultivation, toxinotyping, and ribotyping of positive samples. The toxinotyping of 96 samples revealed that 44.8% of typed strains could produce toxins A and B encoded by tcdA and tcdB, respectively. The ribotyping of the same samples revealed two epidemic peaks, caused by the regionally most prevalent ribotype 176 (n = 30, 31.3). C. difficile infection incidence ranged between 5.5 and 4.2 cases per 10,000 patient-bed days. Molecular diagnostics and molecular epidemiology are the two most developing parts of clinical laboratories. The correct applications of molecular methods help ensure greater safety in hospitals.

• Klíčová slova
• Clonal spreading, Clostridioides difficile, Health care facility, Ribotyping, Toxinotyping,
• MeSH
• bakteriální toxiny * MeSH
• Clostridioides difficile * genetika   MeSH
• Clostridioides MeSH
• klostridiové infekce * diagnóza   epidemiologie   MeSH
• lidé MeSH
• nemocnice univerzitní MeSH
• poskytování zdravotní péče MeSH
• ribotypizace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• bakteriální toxiny * 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.

• Klíčová slova
• MLVA, Slovakia, Thr82Ile, binary toxin, ribotyping, tcdC, Δ117,
• Publikační typ
• časopisecké články MeSH

OBJECTIVES: To investigate the relationship between Clostridium (Clostridioides) difficile strain characteristics and C. difficile infection (CDI) outcome. METHODS: Between October and December 2017, 16 hospitals collected epidemiological data according to the European Centre for Disease Prevention and Control (ECDC) surveillance protocol for CDI. C. difficile isolates were characterized by ribotyping, toxin genes detection and antibiotic susceptibility testing to metronidazole, vancomycin and moxifloxacin. RESULTS: The overall mean CDI incidence density was 4.5 [95% CI 3.6-5.3] cases per 10,000 patient-days. From the 433 CDI cases, 330 (76.2%) were healthcare-associated, 52 (12.0%) cases were community-associated or of unknown origin and 51 (11.8%) CDI cases recurrent; a complicated course of CDI was reported in 65 cases (15.0%). Eighty-eight (20.3%) of patients died and 59 of them within 30 days after the CDI diagnosis. From the 379 C. difficile isolates, the most prevalent PCR ribotypes were 001 (n = 127, 33.5%) and 176 (n = 44, 11.6%). A total of 186 (49.1%) isolates showed a reduced susceptibility to moxifloxacin (> 4 mg/L) and 96.4% of them had Thr82Ile in the GyrA. Nineteen isolates revealed reduced susceptibility to metronidazole and two isolates to vancomycin (> 2 mg/L). A fatal outcome was associated with a reduced susceptibility to moxifloxacin, the advanced age of the patients and a complicated course of CDI (p<0.05). No association between ribotype, binary toxin and a reduced susceptibility to moxifloxacin and complicated course or recurrent CDI was found. CONCLUSIONS: A reduced susceptibility to moxifloxacin, in causative C. difficile strains was associated with fatal outcome of the patients, therefore it is an important marker in surveillance of CDI.

• Klíčová slova
• Clostridioides difficile infection, Clostridium difficile infection, Czech Republic, Mortality, Moxifloxacin, PCR ribotype 001, PCR ribotype 176,
• MeSH
• antibakteriální látky terapeutické užití   MeSH
• Clostridioides difficile účinky léků   MeSH
• feces mikrobiologie   MeSH
• infekce spojené se zdravotní péčí MeSH
• klostridiové infekce farmakoterapie   mikrobiologie   mortalita   MeSH
• lidé MeSH
• mikrobiální testy citlivosti MeSH
• moxifloxacin terapeutické užití   MeSH
• ribotypizace MeSH
• senioři MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• antibakteriální látky MeSH
• moxifloxacin 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 epidemiologie   MeSH
• Názvy látek
• bakteriální proteiny MeSH
• enterotoxiny MeSH
• glutamátdehydrogenasa MeSH

The aim of the study was to provide an update on the epidemiology of C. difficile infection (CDI) in a representative number of hospitals within the Czech Republic in 2015. In 2015, twenty-eight Czech hospitals were invited to participate in a CDI study. Laboratories sent the first 20 consecutive C. difficile isolates for characterization by capillary-electrophoresis (CE) ribotyping and the presence of toxin genes and collected patient data on previous hospitalization, antibiotic treatment, the number of CDI episodes and the course of CDI. The mean incidence of CDI was 5.2 [95% CI 4.2-6.2] cases per 10,000 patient-bed days in 27 hospitals in 2015. Of 490 C. difficile isolates, the prevalent PCR ribotypes (RTs) were 001 (n = 164, 33.5%) and 176 (n = 125, 25.5%) followed by 014 (n = 37, 7.6%), 012 (n = 17, 3.5%), 020 (n = 16, 3.3%), 017 (n = 14, 2.9%) and 002 (n = 11, 2.2%). A severe course of CDI was reported in 104 cases (21.2%) with significant association with RT001 infection (p = 0.03). CDI recurrence was 10.8% (n = 53). The previous use of fluoroquinolones was associated with RTs 001 and 176 CDIs (p = 0.046 and p = 0.041). We observed a persistence of RTs 001 and 176 CDIs in the Czech Republic that was associated with the previous use of fluoroquinolones. This highlights the need for a reduction in fluoroquinolone use in Czech hospital settings.

• Klíčová slova
• CDI surveillance, Capillary electrophoresis ribotyping, Clostridium difficile, Clostridium difficile infection, Fluoroquinolones, Toxin genes,
• MeSH
• antibakteriální látky terapeutické užití   MeSH
• bakteriální léková rezistence fyziologie   MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• fluorochinolony terapeutické užití   MeSH
• infekce spojené se zdravotní péčí epidemiologie   mikrobiologie   MeSH
• lidé MeSH
• nemocnice MeSH
• pseudomembranózní enterokolitida farmakoterapie   epidemiologie   mikrobiologie   MeSH
• ribotypizace MeSH
• senioři MeSH
• Check Tag
• lidé MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• antibakteriální látky MeSH
• fluorochinolony MeSH

In 2011-2012, a survey was performed in three regional hospitals in the Czech Republic to determine the incidence of Clostridium difficile infections (CDIs) and to characterize bacterial isolates. C. difficile isolates were characterized by PCR ribotyping, toxin genes detection, multiple-locus variable-number tandem-repeat analysis (MLVA), and antimicrobial susceptibility testing to fidaxomicin, vancomycin, metronidazole, clindamycin, LFF571, and moxifloxacin using agar dilution method. The incidence of CDI in three studied hospitals was 145, 146, and 24 cases per 100,000 inhabitants in 2011 and 177, 258, and 67 cases per 100,000 inhabitants in 2012. A total of 64 isolates of C. difficile was available for molecular typing and antimicrobial susceptibility testing. 60.9% of the isolates were classified as ribotype 176. All 41 isolates of ribotypes 176 and 078 were positive for the presence of binary toxin genes. Ribotype 176 also carried 18-bp deletion in the regulatory gene tcdC. Tested isolates of C. difficile were fully susceptible to vancomycin and metronidazole, whereas 65.1% of the isolates were resistant to moxifloxacin. MLVA results indicated that isolates from three different hospitals were genetically related, suggesting transmission between healthcare facilities.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální toxiny analýza   genetika   MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• diskové difúzní antimikrobiální testy MeSH
• incidence MeSH
• klostridiové infekce epidemiologie   mikrobiologie   MeSH
• lidé MeSH
• minisatelitní repetice MeSH
• molekulární typizace * MeSH
• nemocnice MeSH
• ribotypizace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika epidemiologie   MeSH
• Názvy látek
• antibakteriální látky MeSH
• bakteriální toxiny 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 epidemiologie   MeSH
• Názvy látek
• antibakteriální látky MeSH