© 2016 Royal College of Pathologists of Australasia. Clostridium difficile rose in prominence in the early 2000s with large-scale outbreaks of a particular binary toxin-positive strain, ribotype 027, in North America and Europe. In Australia outbreaks of the same scale had not and have not been seen. A survey of C. difficile across Australia was performed for 1 month in 2010. A collection of 330 C. difficile isolates from all States and Territories except Victoria and the Northern Territory was amassed. PCR ribotyping revealed a diverse array of strains. Ribotypes 014/020 (30.0%) and 002 (11.8%) were most common, followed by 054 (4.2%), 056 (3.9%), 070 (3.6%) and 005 (3.3%). The collection also contained few binary toxin positive strains, namely 027 (0.9%), 078 (0.3%), 244 (0.3%), 251 (0.3%) and 127 (0.3%). The survey highlights the need for vigilance for emerging strains in Australia, and gives an overview of the molecular epidemiology of C. difficile in Australia prior to an increase in incidence noted from mid-2011.