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

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

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

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

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

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.

• 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

AIM: To obtain standardized epidemiological data for Clostridium difficile infection (CDI) in Slovakia. METHODS: Between October and December 2016, 36 hospitals in Slovakia used the European Centre for Disease Prevention and Control (ECDC) Clostridium difficile infection (CDI) surveillance protocol. RESULTS: The overall mean CDI incidence density was 2.8 (95% confidence interval 1.9-3.9) cases per 10 000 patient-days. Of 332 CDI cases, 273 (84.9%) were healthcare-associated, 45 (15.1%) were community-associated, and 14 (4.2%) were cases of recurrent CDI. A complicated course of CDI was reported in 14.8% of cases (n=51). CDI outcome data were available for 95.5% of cases (n=317). Of the 35 patients (11.1%) who died, 34 did so within 30 days after their CDI diagnosis. Of the 78 isolates obtained from 12 hospitals, 46 belonged to PCR ribotype 001 (59.0%; 11 hospitals) and 23 belonged to ribotype 176 (29.5%; six hospitals). A total of 73 isolates (93.6%) showed reduced susceptibility to moxifloxacin (ribotypes 001 and 176; p< 0.01). A reduced susceptibility to metronidazole was observed in 13 isolates that subsequently proved to be metronidazole-susceptible when, after thawing, they were retested using the agar dilution method. No reduced susceptibility to vancomycin was found. CONCLUSIONS: These results show the emergence of C. difficile ribotypes 027 and 176 with a predominance of ribotype 001 in Slovakia in 2016. Given that an almost homogeneous reduced susceptibility to moxifloxacin was detected in C. difficile isolates, this stresses the importance of reducing fluoroquinolone prescriptions in Slovak healthcare settings.

• MeSH
• antibakteriální látky farmakologie   MeSH
• Clostridioides difficile klasifikace   účinky léků   genetika   izolace a purifikace   MeSH
• incidence MeSH
• klostridiové infekce epidemiologie   mikrobiologie   MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• moxifloxacin farmakologie   MeSH
• ribotypizace MeSH
• senioři MeSH
• Check Tag
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Slovenská republika MeSH

Clostridioides difficile infection (CDI) remains a major healthcare problem worldwide, however, little is known about CDI epidemiology in Iran. Between December 2004 and November 2018, 3649 stool samples were collected from patients in 69 hospitals and medical centres in Tehran and were cultured for the presence of C. difficile; isolates were characterized by PCR ribotyping and toxin genes detection. A total of 582 C. difficile isolates were obtained and the overall CDI prevalence was 15.9%; 290 (49.8%) cases were healthcare-associated (HA) and 292 (50.2%) cases were community-associated (CA). Of these, DNA of 513 isolates submitted for ribotyping. The ribotype and/or WEBRIBO type could be assessed in 366 (62.9%) isolates. The most frequent RTs were 001 (n = 75, 12.9%), 126 (n = 65, 11.2%) and 084 (n = 19, 3.3%); the toxin gene profile tcdA+B+/cdtA+B+ (n = 112, 19.2%) was the most common. Fifteen C. difficile isolates (2.6%) did not carry any toxin genes. There was no difference between frequently found RTs in HA-CDI and CA-CDI, except for RT 029 which was more likely to be associated with healthcare origin (12/15, p-value = 0.02). No isolate of RTs 027 or 078 was identified. Importantly, RTs 031, 038, 039, 084, 085 reported previously as RTs with an absence of toxin genes, revealed the presence of toxin genes in our study. Using Simpson's reciprocal index of diversity, we found that RT diversity decreased as the prevalence of the RT 084 increased (R = -0.78, p-value = 0.041). Different patterns in CDI epidemiology underscore the importance of local surveillance and infection control measures in Tehran healthcare settings.

• MeSH
• Clostridioides difficile klasifikace   genetika   izolace a purifikace   MeSH
• dějiny 21. století MeSH
• dítě MeSH
• dospělí MeSH
• fylogeneze MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• předškolní dítě MeSH
• průřezové studie MeSH
• pseudomembranózní enterokolitida dějiny   mikrobiologie   MeSH
• ribotypizace MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dějiny 21. století MeSH
• dítě MeSH
• dospělí MeSH
• kojenec 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
• historické články MeSH
• Geografické názvy
• Írán MeSH

This study aimed to characterize Clostridium difficile isolates cultured from stool samples of patients with C. difficile infection (CDI) and swabs from a medical environment in a gastroenterology center in Tehran, Iran. A total of 158 samples (105 stool samples from hospitalized patients and 53 swabs from medical devices and the environment) were collected from January 2011 to August 2011 and investigated for the presence of C. difficile by direct anaerobic culture on a selective media for C. difficile. C. difficile isolates were further characterized by capillary electrophoresis (CE) ribotyping and toxin gene multiplex PCR. Of 158 samples, C. difficile was cultured in 19 of 105 stool samples (18%) and in 4 of 53 swabs (7.5%). C. difficile PCR ribotype (RT) 126 was the most common RT in the study (21.7%). Further RTs were: 001, 003, 014, 017, 029, 039, 081, 103 and 150. RTs 126, 001, 150 were cultured from both the stool samples and swabs of medical devices and the hospital environment which suggest a possible route of transmission.

• MeSH
• bakteriální toxiny genetika   MeSH
• Clostridioides difficile klasifikace   genetika   růst a vývoj   izolace a purifikace   MeSH
• feces mikrobiologie   MeSH
• genetická variace * MeSH
• lidé MeSH
• nemocnice MeSH
• polymerázová řetězová reakce MeSH
• ribotypizace * MeSH
• zdravotnické prostředky mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Írán 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