Background: Timely, accurate and accessible antimicrobial susceptibility data are essential for effective clinical and public health responses. In northern Australia, bacterial infection rates are high but resistance to antibiotics is poorly described. HOTspots (amrhotspots.com.au) is a regional, laboratory-based, passive surveillance system that aims to inform clinical management of patients, treatment guidelines and policies. Here we evaluate the utility of this prototype system to end users. Methods and materials: The evaluation was guided by the Centre for Disease Control's framework for evaluating public health surveillance systems. Evidence to assess the system attributes of simplicity, flexibility, acceptability, timeliness, and stability was collected through interviews with clinicians, guideline contributors and policy-makers. Data quality and sensitivity were determined by interviews and an audit of HOTspots data against available antimicrobial susceptibility data. Representativeness was analysed by comparing population coverage of participating and non-participating pathology service providers. Results: The system is simple in structure and operation but could be improved by standardising data collection from laboratories. The data management process facilitates system flexibility, but platform limitations are a barrier to the simplicity, flexibility and acceptability of the system. Acceptability was reduced by the inability to support nuanced clinical and public health decision-making, although enhanced by timely data provision. Future challenges to stability include sustainable governance, funding and human resourcing. Data completeness was high and validity good, though could be improved through quality assurance. The system was found to be sensitive, with an average 94.9% agreement between HOTspots data and local susceptibility data. Three major pathology service providers, who service public and private, hospital and community populations, have contributed data, establishing high quality representativeness of the system. Conclusion: HOTspots’ strength lies in its provision of timely, representative and accurate antimicrobial susceptibility data to end users in northern Australia. Recommendations to enhance the system include standardising data collection, upgrading the platform and working with end users to build more nuanced clinical and public health decision-making support. Sustainable governance and resourcing are essential to achieving this aim. Successful implementation of HOTspots into regional surveillance activities will innovate the way health services receive and act on changing patterns of antimicrobial resistance.