Summary Background: Proton pump inhibitors (PPIs) and H2 receptor antagonists (H2RAs) may play an important role on the onset of Clostridium difficile-associated disease (CDAD) in adults. The impact of Clostridium difficile on children treated with gastric acid-suppressing agents is unknown. Aim: To investigate the relationship between CDAD and exposure to acid suppressive therapy in hospitalized pediatric patients. Patients and Methods: We reviewed the medical records of children with a diagnosis of protracted diarrhea and abdominal pain, whose stool was analyzed for C. difficile toxins. We identified 68 patients with CDAD. For each case patient, we randomly selected one control subjects with stool analysis negative for C. difficile. Comorbid illnesses, previous hospitalizations, antibiotics, corticosteroids, immunosuppressants and gastric acid suppressing exposures were recorded. Results: The use of PPI was significantly higher in C. difficile positive group compared with C. difficile negative group [odds ratio (OR): = 4.5; 95% confidence interval (CI) = 1.4 to 14.4]. We also found a trend for the use of H(2)RAs in patients infected by C. difficile respect to C. difficile negative comparison groups (OR: = 3.8; 95% CI = 0.7 to 18.9). Conclusions: Children exposed to PPIs therapy seem to be at higher risk for the development of CDAD.