Abstract: A total of 29 Staphylococcus aureus strains isolated from human, animals and their environment were subjected to genotypic analysis on the basis of coagulase gene polymorphism. The coagulase gene was amplified using a pair of oligonucleotide nested primers. Presumptive phenotypic identification of the strains showed production of free and bound coagulases, production of acetion, anaerobic utilization of mannitol with acid production. PCR-amplified coagulase genes of S. aureus revealed different pattern in base pair lengths and number of amplified bands. There were no obvious specific PCR pattern for all types of isolates thus, genotypic clustering correlated to human, animal and environmental isolates was not passable. Given the specificity of the coagulase gene, the isolates were thus confirmed to be belong to the coagulase positive S. aurueus. Out of 29 PCR-amplification isolates, 21 produced a single band while 8 isolates produced two bands. The length of the amplicon ranged from 430 to 1000 bp. Amplicons of the 21 isolates were thus categorized as 670, 930, 950 and 1000 bp. In conclusion, Amplification of coagulase gene is useful in confirmation of coagulase gene positive S. aurueus.