ObjectivesTo assess changes in the frequency of vitamin D testing and detection of moderate/severe vitamin D deficiency (<30 nmol/L) among adults after the introduction of new Medicare Benefits Schedule (MBS) rebate criteria (November 2014), and their relationship to sociodemographic and clinical characteristics.DesignDynamic (open) cohort studySettingPrimary careParticipantsAbout 1.5 million ‘active’ patients aged 18+ years visiting a general practitioner and included in the National Prescribing Service MedicineInsight database.Outcome measuresThe frequency of vitamin D testing (per 1000 consultations) and moderate/severe vitamin D deficiency (%) recorded between October 2013 and March 2016, stratified by the release of the new MBS criteria for rebate.ResultsMore patients were female (57.7%) and 30.2% were aged 60+ years. Vitamin D testing decreased 47% (from 40.3 to 21.4 tests per 1000 consultations) after the new MBS criteria, while the proportion of tests with no indication for being performed increased from 71.3% to 76.5%. The proportion of patients identified as moderate/severe vitamin D deficient among those tested increased from 5.4% to 6.5%. Practices located in high socioeconomic areas continued to have the highest rates of testing, but moderate/severe vitamin D deficiency detection remained 90% more frequent in practices from low socioeconomic areas after the rebate change. Furthermore, the frequency of individuals being tested was reduced independent of the patients’ sociodemographic or clinical condition, and the gap in the prevalence of vitamin D deficiency detection between those meeting or not meeting the criteria for being tested remained the same. Moderate/severe vitamin D deficiency detection decreased slightly among patients with hyperparathyroidism or chronic renal failure.ConclusionsAlthough the new criteria for rebate almost halved the frequency of vitamin D testing, it also lessened the frequency of testing among those at higher risk of deficiency, with only a small improvement in vitamin D deficiency detection.