ArticleAntimicrobial resistance and infection control2021
Simultaneous detection and ribotyping of Clostridioides difficile, and toxin gene detection directly on fecal samples.
Article in Antimicrobial resistance and infection control, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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7 citing papers in PubMed, 9 citations in OpenAlex.
- Fecal microbiota composition is a better predictor of recurrent Clostridioides difficile infection than clinical factors in a prospective, multicentre cohort study.BMC infectious diseases · 2024Observational
- Usefulness of Capillary Gel Electrophoresis-Based PCR for Detection ofGels (Basel, Switzerland) · 2024Article
- Comparison of toxin gene expression levels and molecular typing ofGastroenterology and hepatology from bed to bench · 2024Article
- Insights into the Evolving Epidemiology ofAntibiotics (Basel, Switzerland) · 2023Review
- Prevalence of diagnostically-discrepantFrontiers in medicine · 2023Article
- Detection and Genomic Characterisation ofPathogens (Basel, Switzerland) · 2022Article
- Comparative Evaluation of Three Immunoassays for the Simultaneous Detection ofMicroorganisms · 2022Article
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Authors and funding
9 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundClostridioides difficile is the most common cause of nosocomial diarrhea. Ribotyping of cultured strains by a PCR-based test is used to study potential transmission between patients. We aimed to develop a rapid test that can be applied directly on fecal samples for simultaneous detection and ribotyping of C. difficile, as well as detection of toxin genes.
methodsWe developed a highly specific and sensitive primer set for simultaneous detection and ribotyping of C. difficile directly on total fecal DNA. Toxin genes were detected with primers adapted from Persson et al. (Clin Microbiol Infect 14(11):1057-1064). Our study set comprised 130 fecal samples: 65 samples with positive qPCR for C. difficile toxin A/B genes and 65 C. difficile qPCR negative samples. PCR products were analyzed by capillary gel electrophoresis.
resultsRibosomal DNA fragment peak profiles and toxin genes were detected in all 65 C. difficile positive fecal samples and in none of the 65 C. difficile negative samples. The 65 samples were assigned to 27 ribotypes by the Dutch reference laboratory. Our peak profiles corresponded to these ribotypes, except for two samples. During a C. difficile outbreak, patients were correctly allocated to the outbreak-cluster based on the results of direct fecal ribotyping, before C. difficile isolates were cultured and conventionally typed.
conclusionC. difficile ribotyping directly on fecal DNA is feasible, with sensitivity and specificity comparable to that of diagnostic toxin gene qPCR and with ribotype assignment similar to that obtained by conventional typing on DNA from cultured isolates. This supports simultaneous diagnosis and typing to recognize an outbreak.
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