Antibiotic resistance has emerged as one of the most critical global public health challenges, threatening the effective prevention and treatment of a wide range of infectious diseases. The increasing prevalence of resistant pathogens in hospital settings significantly complicates clinical management, prolongs hospital stays, raises healthcare costs, and increases morbidity and mortality rates. This research paper investigates the prevalence and distribution of antibiotic resistance patterns among clinical isolates obtained from hospital laboratories. The study focuses on commonly isolated bacterial pathogens from various clinical samples, including blood, urine, sputum, pus, and wound swabs. Standard microbiological techniques were employed for isolation and identification of bacterial strains, followed by antimicrobial susceptibility testing using the Kirby–Bauer disk diffusion method in accordance with Clinical and Laboratory Standards Institute (CLSI) guidelines. The findings reveal a high prevalence of multidrug-resistant organisms, particularly among Escherichia coli, Staphylococcus aureus, Klebsiella pneumoniae, and Pseudomonas aeruginosa. Resistance to commonly prescribed antibiotics such as penicillins, cephalosporins, and fluoroquinolones was notably high, while comparatively lower resistance was observed against carbapenems and glycopeptides. The study underscores the urgent need for continuous surveillance of antimicrobial resistance, rational antibiotic prescribing practices, and strengthened infection control measures within hospital environments. The outcomes of this research contribute valuable insights for clinicians, microbiologists, and policymakers in designing effective antimicrobial stewardship programs aimed at curbing the growing threat of antibiotic resistance.