The phenomenon of multidrug resistance (MDR) in bacteria represents a significant global health challenge, particularly in hospital environments where resistance genes proliferate through mechanisms such as class 1 integrons and ISCR1 elements. This study investigates the impact of these genetic elements in facilitating antibiotic resistance genes (ARG) dissemination among non-pathogens and key pathogens, specifically Pseudomonas aeruginosa and Staphylococcus aureus, which are notorious for causing nosocomial infections. Through isolation and identification of bacterial strains, antibiotic susceptibility testing, and genomic analysis, we demonstrate a concerning prevalence of resistance to critical antibiotics, particularly β-lactams, aminoglycosides, and fluoroquinolones. Our findings reveal that both Int1 and ISCR1 contribute to the genetic mobility of resistance traits, complicating treatment protocols and elevating healthcare costs due to ineffective therapies. The study emphasizes the urgent need for continued research into the mechanisms of antibiotic resistance dissemination and the development of novel antimicrobial strategies targeting these genetic elements. Furthermore, our results highlight the importance of understanding the historical context of antibiotic resistance gene mobilization as a means to inform healthcare practices and mitigate the rising threat of MDR bacteria on a global scale.