The emergence of Clostridium difficile PCR-ribotype 001 in Slovakia.

Objective: 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.