Background: Antimicrobial resistance (AMR) poses a major global health challenge, leading to treatment failure, prolonged hospital stay, and increased mortality. Local hospital antibiograms are essential tools for guiding empirical therapy and strengthening antimicrobial stewardship. Objectives: To determine the distribution of clinical isolates and assess antibiotic resistance patterns over one year in a tertiary care hospital. Materials and Methods: This cross-sectional laboratory-based study was conducted from September 2024 to August 2025 at a 1500-bedded tertiary care hospital in Central India. All clinical samples from OPD, IPD, and ICUs were processed using standard microbiological methods. Only the first isolate per patient was included. Identification and antimicrobial susceptibility testing were performed using the VITEK 2 system, interpreted as per CLSI 2024 guidelines, and analyzed using WHONET 2024. Results: Of 1,065 culture-positive samples, 76% were Gram-negative bacteria, with Escherichia coli being the most common isolate, followed by Klebsiella pneumoniae and Pseudomonas aeruginosa. High resistance was observed among Gram-negative organisms, while fosfomycin and nitrofurantoin remained effective against urinary E. coli. MRSA prevalence was 58%, with linezolid and vancomycin showing high efficacy. Conclusion: The study demonstrates a high burden of multidrug-resistant pathogens, emphasizing the need for regular antibiogram-based surveillance to support rational antibiotic use.