The emergence of Clostridium difficile PCR-ribotype 001 in Slovakia

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.

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.

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.

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.