The incidence and disease severity of Clostridium difficile infection are rising. There is increasing evidence of a potential association between proton‐pump inhibitors (PPI) and C. difficile infection. We performed a case–control study to examine the relationship between PPI and polymerase chain reaction (PCR)‐proven C. difficile infection in 137 hospitalised patients in a tertiary hospital in Western Australia. Only antibiotic exposure within 3 months prior to onset of diarrhoea was associated with PCR‐proven C. difficile infection (odds ratio 5.97, 95% confidence interval 2.40–14.8, P= 0.001). A restricted analysis on those who had exposure to antibiotics within 3 months before the onset of diarrhoea did not change the negative association between PPI exposure and PCR‐proven C. difficile infection. Long‐term PPI usage and intensity of PPI exposure prior to onset of diarrhoea were not significantly associated with C. difficile infection.