Antibiotic-Associated Diarrhea Mediated by Toxigenic Clostridioides Difficile Infection in Iran: A Clinical Survey by Multiplex Real-Time Polymerase Chain Reaction and Toxigenic Culture

Sepideh Khodaparast, Ashraf Mohabati Mobarez, Nima Khoramabadi, Mohammad Vasei, Mohammad Kamalabadi-Farahani, Mehdi Saberifiroozi



Clostridioides difficile is one of the important causes of hospital infections worldwide.

Regarding the importance of Clostridioides difficile infection (CDI) epidemics and limited available prevalence reports of CDI in Iran, the present investigation was done on the incidence of CDI in hospitalized patients from 2017-2018.

Our study outlined the requirements for CDI and followed up on the causative agent of antibiotic-associated diarrhea (AAD).

Materials and Methods: We evaluated the CDI rates by multiplex real-time polymerase chain reaction (PCR) assay directly from inpatient fecal samples with a history of antibiotic therapy (2-8 weeks) and combined with anaerobic culture and toxicity assessments to isolate toxigenic and non-toxigenic types.

The results were analyzed through one-way analysis of variance (ANOVA), pairwise two-tailed correlation, and regression using SPSS software version 25.0 (IBM® SPSS® Statistics, USA).


Among 491 fecal samples, 49 (9.9%) toxigenic C. difficile were characterized by real-time PCR, while 40 were isolated by toxigenic culture and cytotoxicity assay. Toxin profiling showed 43 (9.7%) tcdA+/tcdB+ and 6 (1.4%) tcdB+.


 A considerable prevalence of CDI among patients with AAD was demonstrated, and causative organisms mostly produce both toxins A and B. Since C. difficile still has problematic treatment and is costly, rapid and early detection may help to curb C. difficile infection more efficaciously.


Antibiotic-associated diarrhea; Clostridioides difficile; Real-time PCR; tcdA; tcdB

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