Objectives: The aim of this study was to describe oral mucosal screening and referral attitudes of Australian oral health therapists (OHTs) and dental hygienists (DHs). Methods: Questionnaires were distributed to participants who attended dental hygiene courses run in both regional and metropolitan Queensland. Results: One hundred and two participants comprised 58 OHTs and 44 DHs, with a mean of 8.9 years since graduation. Thirty-four participants worked in regional locations, while 68 were from metropolitan areas. 97% of participants agreed that mucosal screening should be performed for all new and recall patients, while a minority (5%) agreed that patients will detect an oral mucosal change themselves. The majority (77%) agreed that oral cancer would be encountered in their practising career. Most participants (81%) felt comfortable discussing the presence of a suspicious lesion with patients and 88% agreed that it was their role to screen. In terms of barriers to oral cancer screening, lack of training was seen as the most prevalent barrier (56%) followed by lack of confidence (51%). Lack of time was seen as the third most prevalent barrier (40%), and lack of financial incentives was the least prevalent barrier (16%). Conclusions: Oral health therapists and DHs understand the importance of oral mucosal screening and are likely to be alert to oral mucosal changes. While lack of time and financial incentives was perceived to be impediments to mucosal screening, lack of confidence and training was the most prevalent barriers. This issue should be addressed through implementation of effective continuing education courses targeting oral cancer screening and referral practices.