Objectives Staphylococci are natural inhabitants of skin, glands and mucous membranes of mammals and birds and play very important role in human infections as well. Development and evaluation of effective and reliable identification tools is needed for routine and rapid identification of increasing number of Staphylococcus spp. occurring in human clinical material. Method A group of 68 type and reference staphylococcal strains representing 22 taxa of human associated staphylococci was analyzed by rep-PCR fingerprinting with the (GTG)5 primer. The clinical isolates as well as strains isolated from healthy hosts and environmental samples were tested. Results All tested strains were typeable by the (GTG)5-PCR producing DNA fragments ranging from 200 bp to 4500 bp. Majority of the tested species revealed species-specific fingerprints.