Background: Pseudomonas aeruginosa is considered as a major cause of hospital-acquired infections due to its high antibacterial resistance. Biofilm formation is a well-known pathogenic mechanism in P. aeruginosa infections, since sessile bacteria are protected in an extracellular matrix of exopolysaccharide. The expression of polysaccharide synthesis locus (pslA gene) can be important for biofilm formation by P. aeruginosa. Objectives: The purpose of this research was to evaluate the antibiotic resistance pattern and distribution of the pslA gene among biofilm-producing P. aeruginosa isolates obtained from waste water of Burn Centre in Guilan, Iran. Materials and Methods: Fifty isolates of P. aeruginosa were obtained from waste water of a burn center. The P. aeruginosa isolates were identified using standard bacteriological procedures. Drug susceptibility test was performed by disk diffusion method for all the isolates against nine antimicrobial agents. Biofilm formation was measured by microtiter plate assay. Polymerase chain reaction (PCR) was used to identify the presence of the pslA gene among the isolates. Results: Biofilm formation was observed in 70 of the P. aeruginosa isolates. The potential formation of biofilm was significantly associated with resistance to gentamicin, imipenem, tobramycin and piperacillin. In addition, the pslA gene only existed in biofilm-producing isolates with a frequency of 42.9 (n = 15).Conclusions: The findings of the present study well demonstrated that the P. aeruginosa biofilm-producing isolates were more resistant to the tested antibiotics. Furthermore, because of wide distribution, it seems that the pslA gene is associated with biofilm formation. � 2015, Ahvaz Jundishapur University of Medical Sciences