title: Laboratory diagnosis of Clostridium difficile associated diarrhoea and molecular characterization of clinical isolates.
abstract: We evaluated a three-step algorithm for laboratory diagnosis of Clostridium difficile-associated diarrhoea (CDAD). First, stool specimens were screened using an EIA test for glutamate dehydrogenase detection. Screen-positive specimens were tested by a rapid cytotoxintoxin A/B assay and subjected to stool culture. All cultures positive for C. difficile underwent toxigenic culture. The results showed that toxigenic culture allowed us to recover 37/156 (24.4%) stool samples harbouring toxigenic C. difficile that would have been missed by using faecal cytotoxin assay alone. This determined an increase in infection prevalence of 4.2% (from 11.4% to 15.6 %). Furthermore, to characterize the clinical Clostridium difficile isolates and the distribution of PCR ribotypes circulating in the San Carlo Borromeo hospital, molecular typing using semi-automated repetitive-sequence-based PCR (rep- PCR) and PCR ribotyping, and an evaluation of the antibiotic resistance were also performed. Among them, 71 indistinguishable strains were detected by rep-PCR and 83 by PCR-ribotyping revealing C. difficile outbreaks in our hospital. A total of 6 different ribotypes were obtained by PCR ribotyping. The most frequent ribotype was 018 (88.2%) that also showed resistance to moxifloxacin. In one case, uncommon PCR ribotype 186 was also identified.
authors: U.O.C. Microbiologia e Virologia, A.O. Ospedale San Carlo Borromeo, Milano, Italy.
authors: Giuseppe
authors: G
authors: Russello
authors: Antonio
authors: A
authors: Russo
authors: Francesca
authors: F
authors: Sisto
authors: Maria Maddalena
authors: MM
authors: Scaltrito
authors: Claudio
authors: C
authors: Farina
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000465:Algorithms; D001372:Aza Compounds / Q000494:pharmacology; D001426:Bacterial Proteins / Q000235:genetics; D001427:Bacterial Toxins / Q000235:genetics; D016360:Clostridioides difficile / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D003015:Clostridium Infections / Q000175:diagnosis* / Q000188:drug therapy / Q000382:microbiology; D003967:Diarrhea / Q000188:drug therapy / Q000382:microbiology*; D024881:Drug Resistance, Bacterial; D004768:Enterotoxins / Q000235:genetics; D005243:Feces / Q000382:microbiology*; D005260:Female; D024841:Fluoroquinolones; D005798:Genes, Bacterial; D005969:Glutamate Dehydrogenase / Q000378:metabolism; D006801:Humans; D007124:Immunoenzyme Techniques; D008297:Male; D008795:Metronidazole / Q000494:pharmacology; D008826:Microbial Sensitivity Tests; D008875:Middle Aged; D000077266:Moxifloxacin; D015995:Prevalence; D011804:Quinolines / Q000494:pharmacology; D021521:Ribotyping / Q000379:methods; D012680:Sensitivity and Specificity; D055815:Young Adult
chemical_list: D001372:Aza Compounds; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D004768:Enterotoxins; D024841:Fluoroquinolones; D011804:Quinolines; C041348:tcdA protein, Clostridium difficile; C031815:toxB protein, Clostridium difficile; D008795:Metronidazole; D005969:Glutamate Dehydrogenase; D000077266:Moxifloxacin