Objective The aim of this study is to investigate how healthcare practitioners use real-time prescription monitoring tools in clinical practice. Methods An online survey was distributed to Australian prescribers and pharmacists who use a real-time prescription monitoring tool. Data were analysed and descriptive statistics summarised participant characteristics and responses. A Chi-squared test was conducted to test the difference between prescribers and pharmacists. Results The majority of participants agreed that real-time prescription monitoring (RTPM) information is useful (92.2%) and the tool is valuable for informing clinical decisions (90.2%); however, just over half reported that they had changed their prescribing or dispensing practices as a result of RTPM information (51.0%), and they employed evidence-based clinical interventions to varying degrees. No statistically significant differences were detected between pharmacists and prescribers and perceptions on tool use. Conclusions This is the first known study to investigate practitioner use of RTPM tools in Australia, and is a starting point for further research. What constitutes 'success' in the clinical application of RTPM tools is yet to be realised.