Antimicrobial resistance (AMR) is an escalating global concern, particularly in developing countries like India. A 1-year prospective study was conducted on AMR in human pathogens from Ahmedabad, India. The study aimed to generate an evidence-based database on the AMR profile of pathogens in this region. The study analysed 2204 organisms isolated from various clinical specimens. WHONET software, a specialized tool for AMR data management and interpretation, was used for data management and analysis. The most frequently isolated pathogens were Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Staphylococcus aureus. These pathogens demonstrated varying resistance rates to different antibiotics. E. coli shows a high prevalence of MDR (57%), with 22% indicating possible XDR and 13% showing possible PDR. K. pneumoniae showed even higher rates of MDR (80%), with 57% indicating possible XDR and 54% possible PDR. S. aureus showed MDR in 51% of the isolates, with 11% showing possible XDR and 1% showing possible PDR. The study also identified some priority pathogens according to the World Health Organization (WHO) criteria based on their resistance to specific antibiotics. The study highlighted the significant prevalence of AMR, particularly MDR, among human pathogens in Ahmedabad, emphasizing the need for effective strategies to combat AMR in clinical settings and public health policies. The study has significant implications for understanding the epidemiology and transmission of AMR in this region, as well as for informing the development of guidelines and interventions for rational antibiotic use and infection control.