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Background <p>Knowledge of antimicrobial resistance patterns and <i>C. difficile</i> ribotypes is necessary for more effective treatment of <i>C. difficile</i> infection (CDI) in healthcare settings.</p> Methods and results <p>A total of 180 stool samples were collected from patients who were referred to colonoscopy departments. The Minimum Inhibitory Concentration (MIC) of vancomycin, clindamycin, ciprofloxacin, and ampicillin / sulbactam against <i>C. difficile</i> isolates was determined by Liofilchem<sup>®</sup> MIC Test Strips. Moreover, the susceptibility of isolates to metronidazole was identified using the Disk Diffusion Method (DDM). The Polymerase Chain Reaction (PCR) assay was used to detect the toxin-encoding genes in <i>C. difficile</i> isolates. PCR ribotyping was performed on the toxin-positive <i>C. difficile</i>. The SYBR Green-based Quantitative Real-Time PCR was used to evaluate the relative expression levels of toxin-encoding genes. Overall, 34 (18.9%) isolates confirmed as <i>C. difficile</i>; from these, 26.5%, 35.3%, 35.3%, and 41.2% were positive for <i>tcdA</i>, <i>tcdB</i>, <i>cdtA</i>, and <i>cdtB</i> genes, respectively. Eight isolates were <i>tcdB</i>+/<i>cdtA</i>+/<i>cdtB</i>+, three isolates were <i>tcdA</i>+/<i>tcdB</i>+, and four isolates were <i>tcdA</i>+/<i>cdtB</i>+. Resistance to metronidazole and vancomycin was shown in 8.8% and 5.9% of isolates, respectively. The most frequent ribotypes were RT012 (38%) and RT001 (19%), respectively. <i>tcdA</i> gene expression was significantly high in ribotypes 001 (8.26 fold), 078 (6.35 fold), and 027 (7.65 fold). Moreover, the <i>tcdB</i> relative expression was significantly high in ribotypes 012 (4.75 fold) and 078 (14.45 fold).</p> Conclusions <p>Increase in the expression levels of toxin-encoding genes in RT027 and RT078 revealed that these ribotypes are hypervirulent and can cause severe infections in patients.</p>

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Antibiotic resistance patterns, toxin profiles, and toxin gene expression levels among Clostridioides difficile PCR ribotypes isolated from patients scheduled for colonoscopy

  • 期刊论文

  • 发表日期：

    2025年12月29日

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  • Sahar Sabour,
  • Taher Azimi,
  • Effat Abbasi Montazeri,
  • Seyed Saeed Seyedian,
  • Zahra Farshadzadeh,
  • Mansour Amin

摘要

Background

Knowledge of antimicrobial resistance patterns and C. difficile ribotypes is necessary for more effective treatment of C. difficile infection (CDI) in healthcare settings.

Methods and results

A total of 180 stool samples were collected from patients who were referred to colonoscopy departments. The Minimum Inhibitory Concentration (MIC) of vancomycin, clindamycin, ciprofloxacin, and ampicillin / sulbactam against C. difficile isolates was determined by Liofilchem® MIC Test Strips. Moreover, the susceptibility of isolates to metronidazole was identified using the Disk Diffusion Method (DDM). The Polymerase Chain Reaction (PCR) assay was used to detect the toxin-encoding genes in C. difficile isolates. PCR ribotyping was performed on the toxin-positive C. difficile. The SYBR Green-based Quantitative Real-Time PCR was used to evaluate the relative expression levels of toxin-encoding genes. Overall, 34 (18.9%) isolates confirmed as C. difficile; from these, 26.5%, 35.3%, 35.3%, and 41.2% were positive for tcdA, tcdB, cdtA, and cdtB genes, respectively. Eight isolates were tcdB+/cdtA+/cdtB+, three isolates were tcdA+/tcdB+, and four isolates were tcdA+/cdtB+. Resistance to metronidazole and vancomycin was shown in 8.8% and 5.9% of isolates, respectively. The most frequent ribotypes were RT012 (38%) and RT001 (19%), respectively. tcdA gene expression was significantly high in ribotypes 001 (8.26 fold), 078 (6.35 fold), and 027 (7.65 fold). Moreover, the tcdB relative expression was significantly high in ribotypes 012 (4.75 fold) and 078 (14.45 fold).

Conclusions

Increase in the expression levels of toxin-encoding genes in RT027 and RT078 revealed that these ribotypes are hypervirulent and can cause severe infections in patients.