Pseudomonas aeruginosa is a ubiquitous bacterium that causes various hospital- acquired and community-acquired infections. It has been reported that the clinical isolates of P. aeruginosa are difficult to treat because of their virulence factors and antibiotics resistances. The aim of present study was to screen the antibiotic resistance patterns and the prevalence of virulence factor genes in a set of Pseudomonas aeruginosa isolated from Ogbomoso, and to determine whether a correlation exists between the prevalence of virulence factors and antibiotic resistance of P. aeruginosa. A total of 100 P. aeruginosa isolates were collected from various types of clinical specimens. Antimicrobial susceptibility testing was performed using the Kirby-bauer method. In addition, PCR assays were used for screening four virulence encoding genes (OPRL, LasB, PLCH and ToxA). The results showed that OPRL (79%) and LasB (62%) were the most frequent virulence genes in P. aeruginosa strains, followed by PLCH (41%) and ToxA (35%). The highest resistance was detected towards Piperacillin (42%) and Tetracycline (42%). Moderate rate of resistance (12-39%) were detected towards the other antibiotics. The virulent factors identified in this study provide valuable information regarding the prevalence of resistance genes of P. aeruginosa isolates in Ogbomoso, Nigeria and their potential impact on treatments that exploit the unique physiology of the pathogen. This will be useful for the health workers to improve infection control measures and to establish a surveillance system. Keywords: antibiotic resistance, virulence genes, Pseudomonas Aeruginosa