Background: Pseudomonas aeruginosa is a ubiquitous gram-negative pathogen with a propensity to cause opportunistic infections in humans. Different strains of the organism could colonise patients heralding a wide spectrum of P. aeruginosa infections in the environment., Objective: To analyse isolates of P. aeruginosa from clinical and environmental samples using the Polymerase Chain Reaction (PCR) to establish strain relatedness., Methods: Fifty-two strains of the organism were isolated from wound swabs, urine, sputum of patients and environmental samples from the hospital environment using standard microbiological techniques and ethical consideration. Genomic DNA of the isolates was amplified with primers AF1 (5'-AGA GTT TGA TCC TGG CTCA-3') and 1541R (5'-AAG GAG GTG ATC CAG CC-3')., Results: At least two bands were observed in all isolates typed and band sizes ranged from 0.07 - 1.5kb. The strains were genetically diverse, displaying profiles of 2 - 6 bands between 0.07 - 1.5kb., Conclusion: The study demonstrates that strain diversity could be discerned between strains of P. aeruginosa, circulating in the environment of Buea, a finding which has important epidemiological and clinical significance bearing in mind that this pathogen is highly incriminated in nosocomial infections with attendant social implications. This therefore calls for more attention in the diagnosis and management of P. aeruginosa infections in the environment of Buea, Cameroon.