Antibiotic resistance continues to undermine treatment options worldwide, particularly in developing countries, where access to effective antibiotics is often delayed while resistant strains proliferate. This retrospective, cross-sectional study evaluated resistance patterns and the pathogens and specimen types most commonly isolated at a tertiary care hospital in Jamaica. Antibiograms from January 2019 to December 2023 were reviewed, and annual resistance patterns were compared using chi-square testing (p < 0.05). Over the five-year period, 3,885 pathogens representing 57 microbial species were isolated from specimens including urine, eye swabs, wound, vaginal swabs, aspirates, semen, catheters, stool, ulcer, throat, sputum, and abscess; urine predominated, accounting for 75.19% (n = 2,728) of isolates. Escherichia coli was the most frequently isolated organism (29.66%), followed by Enterobacter species (16.81%), Proteus mirabilis (7.08%), Enterobacter gergoviae (7.03%), coagulase-negative staphylococci (6.75%), Group D streptococci (5.62%), Pseudomonas aeruginosa (4.49%), and Staphylococcus aureus (3.25%). Susceptibility testing most often involved ampicillin, Augmentin, ciprofloxacin, cefuroxime, nitrofurantoin, gentamicin, and ofloxacin, with resistance emerging against all agents to varying degrees and ampicillin showing the highest rates by a clear margin. These findings raise concern about the extent of antimicrobial resistance in this setting, particularly given the pronounced resistance of E. coli to ampicillin and how frequently this drug is used empirically. As a single-center, retrospective analysis, this study cannot capture nationwide trends, underscoring the need for coordinated, country-wide antibiogram tracking to monitor resistance patterns as they evolve rather than assess them retrospectively at individual institutions.