Antibiotic　resistance　threatens　global　public　health.　Clinical　methods　that　simplify　and　accelerate　resistance　diagnosis　are　urgently　needed.　Here　we　describe　a　function-based　antibiotic　resistance　detection　and　classification　strategy　to　improve　diagnosis.　The　method　identifies　resistance　enzymes　by　directly　measuring　the　thermal　signal　generated　when　an　antibiotic　i　enzymatically　degraded.　A　substrate　specificity　profile　is　created　by　analyzing　a　panel　of　antibiotics.　Here　we　show　proof　of　principle　by　differentiating　two　antibiotic　resistance　enzymes　based　on　their　substrate　specificities　profiles.　The　method　provides　a　fast,　simple,　cost　effective　alternative　for　diagnosing　and　classifying　antibiotic　resistance.　(C)　2016　The　Authors.　Published　by　Elsevier　Ltd.