Molecular, microbiological and clinical characterization of Clostridium difficile isolates from tertiary care hospitals in Colombia.
Clara Lina Salazar, Catalina Reyes, Santiago Atehortua, Patricia Sierra, Margarita María Correa, Daniel Paredes-Sabja, Emma Best, Warren N Fawley, Mark Wilcox, Ángel González
Author Information
Clara Lina Salazar: Research Group in Anaerobic Bacteria (GIBA), School of Microbiology, Universidad de Antioquia, Medellín, Colombia.
Catalina Reyes: Basic and Applied Microbiology Research Group (MICROBA), School of Microbiology, Universidad de Antioquia, Medellín, Colombia.
Santiago Atehortua: San Vicente Fundación University Hospital, Medellín, Colombia.
Patricia Sierra: Clínica León XIII, IPS Universitaria, Universidad de Antioquia, Medellín, Colombia.
Margarita María Correa: Molecular Microbiology Group, School of Microbiology, Universidad de Antioquia, Medellín, Colombia.
Daniel Paredes-Sabja: Microbiota-Host Interactions and Clostridia Research Group, Departamento de Ciencias Biológicas, at Universidad Andres Bello, Santiago, Chile.
Emma Best: Departament of Microbiology, Leeds Teaching Hospitals NHS Trust, Leeds, United Kingdom.
Warren N Fawley: Departament of Microbiology, Leeds Teaching Hospitals NHS Trust, Leeds, United Kingdom.
Mark Wilcox: Departament of Microbiology, Leeds Teaching Hospitals NHS Trust, Leeds, United Kingdom.
Ángel González: Basic and Applied Microbiology Research Group (MICROBA), School of Microbiology, Universidad de Antioquia, Medellín, Colombia. ORCID
In Colombia, the epidemiology and circulating genotypes of Clostridium difficile have not yet been described. Therefore, we molecularly characterized clinical isolates of C.difficile from patients with suspicion of C.difficile infection (CDI) in three tertiary care hospitals. C.difficile was isolated from stool samples by culture, the presence of A/B toxins were detected by enzyme immunoassay, cytotoxicity was tested by cell culture and the antimicrobial susceptibility determined. After DNA extraction, tcdA, tcdB and binary toxin (CDTa/CDTb) genes were detected by PCR, and PCR-ribotyping performed. From a total of 913 stool samples collected during 2013-2014, 775 were included in the study. The frequency of A/B toxins-positive samples was 9.7% (75/775). A total of 143 isolates of C.difficile were recovered from culture, 110 (76.9%) produced cytotoxic effect in cell culture, 100 (69.9%) were tcdA+/tcdB+, 11 (7.7%) tcdA-/tcdB+, 32 (22.4%) tcdA-/tcdB- and 25 (17.5%) CDTa+/CDTb+. From 37 ribotypes identified, ribotypes 591 (20%), 106 (9%) and 002 (7.9%) were the most prevalent; only one isolate corresponded to ribotype 027, four to ribotype 078 and four were new ribotypes (794,795, 804,805). All isolates were susceptible to vancomycin and metronidazole, while 85% and 7.7% were resistant to clindamycin and moxifloxacin, respectively. By multivariate analysis, significant risk factors associated to CDI were, staying in orthopedic service, exposure to third-generation cephalosporins and staying in an ICU before CDI symptoms; moreover, steroids showed to be a protector factor. These results revealed new C. difficile ribotypes and a high diversity profile circulating in Colombia different from those reported in America and European countries.
