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