The interfacial transition zone （ITZ） of asphalt mixture at room temperature was analyzed by micro-scratch test combined with scanning electron microscopy （SEM）. The spatial geometric range of ITZ was deduced， and the fracture toughness and friction coefficient were determined. Three-point bending test and nano-indentation test were used for comparative verication. The results show that the ITZ region of asphalt mixture has clear characteristics， and when the load is 100 mN， it is more suitable for the micro-scratch test. Under the scratch effect， a large fracture surface will appear in the ITZ region， and the thickness fluctuates between 10-30 μm， which is similar to the results of nanoindentation test. ITZ friction coefficient increases gradualy from aggregate to asphalt mortar， and tends to be stable after reaching the maximum value. The fracture toughness value obtaind by micro-scratch test is close to the three-point bending test result， and the three-phase medium of aggregate， ITZ and asphalt mortar can be better identified from the fracture toughness curve， indicating that the preliminary analysis of asphalt mixture ITZ can be easily realized by micro-scratch test.