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Cefoperazone-treated Mouse Model of Clinically-relevant Clostridium difficile Strain R20291
Published on: December 10, 2016
An Outbreak of Clostridium (Clostridioides) difficile Infections within an Acute and Long-Term Care Wards Due to
Elena Novakova1, Nina Kotlebova1, Anezka Gryndlerova2
1Department of Microbiology and Immunology, Comenius University Jessenius Faculty of Medicine, 036 01 Martin, Slovakia.
Abstract:
We aimed to characterize Clostridioides difficile isolates cultured during a six-month single-center study from stool samples of patients with C. difficile infection (CDI) genotyped by the Xpert®C. difficile/Epi assay by polymerase chain reaction (PCR) ribotyping, toxin genes' detection and multi-locus variable number tandem repeats analysis (MLVA). The susceptibility to metronidazole, vancomycin and moxifloxacin was determined by agar dilution. In addition, the presence of Thr82Ile in the GyrA and a single nucleotide deletion at position (Δ117) in the tcdC gene were investigated. Between January 1 and June 30, 2016, of 114 CDIs, 75 cases were genotyped as presumptive PCR ribotype (RT) 027 infections using a commercial assay. C. difficile isolates cultured from presumptive RT027 stool samples belonged to RT176. These isolates carried genes for toxin A (tcdA), B (tcdB), binary (cdtA/B) and had Δ117 in the tcdC gene. Using MLVA, the 71/75 isolates clustered into two clonal complexes (CCs). Of these, 39 isolates (54.9%) were from patients hospitalized in acute care and 32 isolates (45.1%) were isolated from patients hospitalized in the long-term care department. All isolates were susceptible to metronidazole and vancomycin, and 105 isolates were resistant to moxifloxacin (92%) carrying Thr83Ile in the GyrA. An outbreak of RT176 CDIs, suspected as RT027, was recognized in a Slovakian hospital. In order to monitor the emergence and spread of RT027-variants, the identification of a presumptive RT027 CDI should be confirmed at a strain level by PCR ribotyping.
Insights
A study identified Clostridioides difficile (C. difficile) isolates as RT176, not the suspected RT027, in a Slovakian hospital outbreak. This highlights the need for strain-level confirmation to monitor C. difficile variants.
Area of Science:
- Microbiology
- Infectious Diseases
- Genetics
Background:
- Clostridioides difficile infection (CDI) poses a significant healthcare challenge.
- Accurate strain identification is crucial for understanding and controlling CDI outbreaks.
- The Xpert® C. difficile/Epi assay is a commercial tool for presumptive identification.
Purpose of the Study:
- To characterize C. difficile isolates from a suspected RT027 outbreak.
- To investigate genotypic and antimicrobial susceptibility profiles of these isolates.
- To evaluate the accuracy of presumptive identification methods for C. difficile strains.
Main Methods:
- Polymerase chain reaction (PCR) ribotyping, toxin gene detection, and multi-locus variable number tandem repeats analysis (MLVA) were employed.
- Antimicrobial susceptibility testing (metronidazole, vancomycin, moxifloxacin) was performed using agar dilution.
- Specific genetic markers (GyrA Thr82Ile, tcdC Δ117) were investigated.
Main Results:
- 75 presumptive RT027 infections were identified as RT176 by culture and further analysis.
- RT176 isolates possessed toxin genes (tcdA, tcdB, cdtA/B) and the tcdC Δ117 mutation.
- All isolates were susceptible to metronidazole and vancomycin; 92% were resistant to moxifloxacin.
Conclusions:
- A C. difficile RT176 outbreak was misidentified as RT027 by a commercial assay.
- Strain-level confirmation using PCR ribotyping is essential for accurate CDI surveillance.
- Monitoring for emerging C. difficile variants requires precise genotypic characterization.
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