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Related Concept Videos

Modern Molecular Taxonomy01:29

Modern Molecular Taxonomy

505
Advancements in molecular biology have revolutionized the identification and characterization of bacteria, with multiple methods leveraging DNA sequencing for enhanced precision. As sequencing technologies improve and costs decline, these approaches are increasingly used in clinical, environmental, and evolutionary studies.Multilocus Sequence Typing (MLST) examines several housekeeping genes, essential chromosomal genes encoding cellular functions, to distinguish strains. Approximately...
505

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Related Experiment Video

Updated: Dec 27, 2025

A Protocol to Characterize the Morphological Changes of Clostridium difficile in Response to Antibiotic Treatment
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Ribotypes in isolates and testing algorithm of C. difficile infections in the studied sample.

Z Stofkova, E Novakova, M Novak

    Bratislavske Lekarske Listy
    |March 3, 2020
    PubMed
    Summary

    Clostridium difficile (C. diff) infections are common in hospitals. This study evaluated a testing algorithm to identify hypervirulent C. diff strains, finding PCR effective for rapid diagnosis and strain identification.

    Keywords:
    diagnosticsreal-time PCR ribotyping.toxigenic C. difficile

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    Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291
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    Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291

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    Area of Science:

    • Clinical microbiology
    • Infectious diseases
    • Molecular diagnostics

    Background:

    • Clostridium difficile is a leading cause of healthcare-associated diarrhea.
    • Identifying hypervirulent strains, such as RT 027, is crucial for patient management.
    • Antibiotic-associated diarrhea necessitates effective diagnostic strategies.

    Purpose of the Study:

    • To evaluate a two-step diagnostic algorithm for Clostridium difficile infection (CDI).
    • To identify hypervirulent Clostridium difficile strains, specifically RT 027.
    • To assess the utility of real-time PCR in CDI diagnosis.

    Main Methods:

    • Retrospective analysis of 1006 patient samples suspected of CDI.
    • Utilized a two-step algorithm involving GDH, toxin A/B immunoassay, and culture.
    • Employed real-time PCR as a confirmatory or third-step diagnostic method.

    Main Results:

    • 202 out of 1006 samples tested positive for CDI using the two-step algorithm.
    • Real-time PCR identified 59 toxigenic and non-RT 027 ribotypes among 64 discordant isolates.
    • No statistically significant association was found between diagnostic parameters, hospitalization length, and C. diff RT027 ribotypes.

    Conclusions:

    • PCR ribotyping revealed a high prevalence of hypervirulent and toxigenic strains.
    • Real-time PCR facilitates rapid laboratory diagnosis and treatment of high-risk patients.
    • Vancomycin resistance was detected in one isolate, highlighting the need for ongoing surveillance.