Background: Infections caused by Pseudomonas aeruginosa raise an important issue in burn patients. Molecular epidemiologic studies have been used for investigating the genetic features of P. aeruginosa and rep-PCR technique has been introduced as a rapid low cost method. Objectives: This study focused on investigating the genetic similarity and antibiotic resistance pattern of P. aeruginosa isolated from the clinical samples of burn patients in a major burn center in Khuzestan Province, Iran. Methods: In a cross sectional study, a total of 75 strains of P. aeruginosa were isolated from burn patients at Taleghani hospital, which is the main burn center in Ahvaz, Iran, during May-September, 2015. Antimicrobial susceptibility of the isolates was detected using the disk diffusion method. Genetic relatedness of the isolates was analyzed by the rep-PCR technique. Results: Antimicrobial susceptibility testing showed more than 80% of P. aeruginosa isolates were resistant to ceftriaxone, cefotaxime, meropenem, piperacillin/tazobactam, ticarcillin, ciprofloxacin, and amikacin. Based on the rep-PCR analysis, 20 different common types and 20 unique patterns were illustrated among P. aeruginosa isolates. Conclusions: According to the findings of our study, there were diverse and high-level resistant P. aeruginosa strains in the major burn center in Khuzestan. Therefore, we faced troubles controlling the diverse P. aeruginosa clones in the burn patients.