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Because Staphylococcus aureus (S. aureus) has variable number of short sequence repeat region in coagulase gene, it has been used to investigate the relatedness of S. aureus isolates. In this study, we isolated S. aureus strains from 20 dairy farms with bovine mastitis from September 2000 to August 2001. PCR-RFLP analysis of coagulase gene revealed 10 different patterns. Most of the S. aureus isolates showed only one coagulase gene RFLP pattern per farm. However, there were several S. aureus clones spreading between dairy farms. All the farms showed poor management conditions of milking machine and milker, indicating that managements for mastitis control program include use of proper milking matching, premilking sanitation, and segregation in the S. aureus infection herd. Our data suggest that PCR-RFLP analysis of coagulase gene might be applicable for the epidemiological investigations of S. aureus isolated from bovine mastitis cows.