Aims: The aim of this study was to investigate plasmid profiles and antibiotic resistance patterns of multidrug resistant Pseudomonas aeruginosa strains isolated from different clinical specimens in Jordan. Methodology and results: Pseudomonas aeruginosa strains were isolated from different clinical specimens from different hospitals and primary health care centers. The antimicrobial susceptibility of P. aeruginosa isolates was determined using the disc diffusion method against 16 commonly used antimicrobial drugs. Plasmid DNAs were extracted from lysed P. aeruginosa cells using the plasmid alkaline lysis method and visualized using agarose gel electrophoresis followed by ethidium bromide staining. The isolated strains of P. aeruginosa were resistant to cefepime (90%), meropenem (70%), ceftazidime (60%), piperacillin (55%), aztreonam (50%), ciprofloxacin and tobramycin (35%), gentamicin (29%), imipenem and amikacin (20%). All the isolates were sensitive to colistin. Plasmid analysis of the clinical isolates showed the presence of 0 to 3 plasmids with a size range of 1 to 25 kb compared to the standard strain of Escherichia coli (ATCC 25922). Conclusion, significance and impact of study: The results obtained in this study showed some correlation between the patterns of antibiotic resistance and plasmid profiles.