DNA analysis enables genome fingerprinting with consequent identification of different individuals. In the agro-food industry, this can have interesting applications for the identification of species and cultivars of both raw materials and processed food. In this investigation, the efficiency of DNA microsatellite analysis in identifying virgin olive oils from different cultivars was evaluated. Ten virgin oils were obtained in the laboratory from olives of 10 different cultivars and the DNA extracted from the cell residues, recovered by oil centrifugation, was used as a template with seven different primer pairs of microsatellite sequences. The electrophoretic patterns showed an adequate level of amplification and were identical to those obtained from leaves and drupes of the same cultivar. By analyzing all the patterns obtained, the smallest number of microsatellites able to distinguish the examined oils was established and an identification key for the different oils was developed.