The Pseudomonas aeruginosa bacterium, which is considered a nosocomial infectious agent, changes its structure due to exposure to various antibiotics and becomes relatively resistant to them, which makes it difficult to treat diseases caused by this bacterium. This study can identify the main genes that make biofilm and allow researchers to inhibit the rate of biofilm formation and spread by making antibiotics that affect these genes, and ultimately improve the patient by inhibiting bacterial growth. In this study, clinical isolates were collected from Afzalipour Hospital in Kerman, Iran and identified as phenotypic and then genotypic. Fifty-five clinical isolates of P. aeruginosa were confirmed. Antibiogram test was performed for meropenem, amikacin, ampicillin–sulbactam, cefotaxime, levofloxacin, rifampin, and tigecycline antibiotics. The ability to form biofilms of isolates was determined by microtiter plate and staining using crystal violet and then spectrophotometry in OD 490 nm. PCR was performed to identify the frequency of pslA and pelB genes. The data showed that the highest age group involved 1 to 15 years included 19% and the lowest age group 26 to 35 years included 5%. The frequency of pslA and pelB genes was 34.5% and 65.5%, respectively, and the range of drug resistance for the mentioned antibiotics was between 72 and 100%. Since the frequency of the pelB gene is about twice as high as that of pslA, it may be concluded that in most of the isolates studied, Psl often has a disorder or that the intracellular level of c-di-GMP has increased sharply.