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Introduction: Cytolethal distending toxin (Cdt) is one of the bacterial toxins that present in a variety of gram-negative human pathogens, such as E. coli, Salmonella spp., and Campylobacter spp. CDT is composed of three subunits encoded by three adjacent genes, including cdtA, cdtB, and cdtC. cdtB has been shown to have toxic activity and cause DNA damage in host cells. Despite its presence in different bacterial species, the role of CdtB in acute and chronic infections, such as gastroenteritis and irritable bowel syndrome (IBS), is unclear. To analyze this correlation, we studied the prevalence of cdtB among different enteropathogenic bacteria in patients with gastroenteritis and IBS compared with healthy people., Materials and Methods: In this cross-sectional descriptive study, 230 stool samples were collected from patients with gastroenteritis, IBS, and healthy people. The presence of CdtB encoding bacteria, including Escherichia coli, Campylobacter spp., Yersinia entercolitica, Providencia alkalifacience, and Salmonella enterica, was examined by polymerase chain reaction using genus-specific primers., Results: Out of 230 stool samples, CdtB encoding Campylobacter spp. were found in 34.6% (52/150), 6.25% (5/80), and 4% (2/50) of the patients with gastroenteritis, IBS, and the control group, respectively. Carriage of CdtB encoding Salmonella enterica was characterized among 5.3% (8/150) of the patients with gastroenteritis and 17.5% (14/80) of the IBS patients. Although none of the patients carried CdtB encoding E. coli and Providencia spp., cdtB of Y. enterocolitica was detected in one of the patients with gastroenteritis (0.6%). Statistical analysis showed significant correlation between infection with CdtB encoding Campylobacter spp. and IBS-D subtype. No significant correlation was found between infection with CdtB encoding bacteria and other clinical and demographic data., Conclusion: Our results confirmed a relatively higher frequency of CdtB encoding bacteria in the intestine of patients with gastroenteritis and those with IBS compared with healthy individuals. Regarding the frequency of CdtB encoding Salmonella and Campylobacter bacteria, it was proposed that infection with these enteropathogens could be considered a risk factor for the development or progression of IBS among Iranian patients. Further studies are needed to establish this involvement., (© 2022. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)