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Background A priority area in the 2016 Victorian Hepatitis B Strategy is to increase diagnostic testing. This study describes hepatitis B testing and positivity trends in Victoria between 2011-2016 using data from a national laboratory sentinel surveillance system. Methods Line-listed diagnostic and monitoring hepatitis B testing data among Victorian individuals were collated from six laboratories participating in the Australian Collaboration for Coordinated Enhanced Sentinel Surveillance (ACCESS) of STIs and blood borne viruses. Diagnostic tests included hepatitis B surface antigen (HBsAg)-only tests and guideline-based hepatitis B tests (defined as a single test event for HBsAg, hepatitis B surface antibody and hepatitis B core antibody). Using available data, the outcome of testing and/or infection were further classified. Measures reported include the total number of HBsAg and guideline-based tests conducted and the proportion positive, classified as either HBsAg positive or chronic hepatitis B infection. Findings The number of HBsAg tests decreased slightly each year between 2011 and 2016 (from n=91,043 to n=79,664 in 2016, p<0.001) whilst the number of guideline-based hepatitis B tests increased (from n=8,732 in 2011 to n=16,085 in 2016, p<0.001). The proportion of individuals classified as having chronic infection decreased from 25% (2011) to 7% (2016) while the proportion classified as susceptible and immune due to vaccination increased from 29% to 39%, and 27% to 34%, respectively (p<0.001). Conclusions Findings indicate an increased uptake of guideline-based hepatitis B testing. The ongoing collection of testing data can help to monitor progress towards implementation of the Victorian Hepatitis B Strategy.