The ghost bat (Macroderma gigas) is listed as Vulnerable in Australia, and is a difficult species to study in the wild. The published literature available on even the most basic aspects of its ecology is limited. This study describes an investigation into the diet of ghost bats occupying the Pilbara region of Western Australia, using identification of dried food remains recovered from beneath roosts in the 1980s and 1990s, and DNA metabarcoding of faecal pellets collected from roost sites during 2011–12. Ghost bat diet in the Pilbara region consists primarily of small mammal and bird species, with a lesser contribution from reptiles (geckoes and skinks) and amphibians. In total, 46 vertebrate taxa were identified, with 32 taxa identified from the dried food remains, and 21 taxa by DNA metabarcoding analysis of the faecal pellets. Only seven of the taxa identified were common to both collection methods, and 32 of those taxa identified represent new prey records for ghost bats in Western Australia, and 19 prey species that had not previously been reported from any other part of Australia. Knowledge of the diet of the ghost bat will provide land managers with important information necessary to ensure the continued survival of this species across its range.