CARRIAGE OF CLOSTRIDIUM DIFFICILE IN HOSPITAL PATIENTS IN SPAIN, INCLUDING MOLECULAR CHARACTERIZATION AND ANTIMICROBIAL SUSCEPTIBILITY OF THE ISOLATES

Increasing age, several co-morbidities, environmental contamination, antibiotic exposure and other intestinal perturbations appear to be the greatest risk factors for C. difficile infection (CDI). Therefore, hospitalized patients are considered particularly vulnerable to CDI. The main objective of this study was to evaluate the prevalence of C. difficile in a Spanish hospital and to characterize the isolates with respect to the PCR-ribotype, antibiotic resistance and toxin activity. In addition, multilocus sequence typing (MLST) was performed in order to study clonal relationships between C. difficile isolates obtained from two different countries.
Culture of samples was performed in a selective medium cycloserine cefoxitin fructose cholate. An identification of the isolated colonies was done by PCR detection of tpi, tcdA, tcdB and cdtA genes. Toxic activity was confirmed by a cytotoxic immunoassay. Further characterization was performed by PCR ribotyping. MLST was used in order to determine genetic relationships between Spanish and Belgian C. difficile isolates recovered from hospital patients in both countries.
C. difficile was frequently detected in hospitalized patients. The isolates belonged to different PCR ribotypes, including type 027. Most of the strains obtained harboured tcdA and tcdB genes. The number of positive faecal samples considerably increased among elderly patients over 65 years old. The multi-locus sequence typing (MLST) analysis showed that some Spanish and Belgian isolates cluster in the same lineage.
This study reveals the circulation of toxigenic C. difficile in a Spanish hospital. The relatedness between Belgian and Spanish isolates indicate a common source or a lack of diversity from some PCR-ribotypes. Moreover, data obtained suggest that the combination of ribotyping and MLST is a good tool for the inter-laboratory comparison of strains among different countries. Continuous inter-country surveillance is suitable to understand the spread of C. difficile isolates among hospitalized patients.