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The study investigated the antibacterial activity of essential oil from the peel of Citrus aurantifolia against eleven multidrug-resistant (MDR) bacterial isolates of clinical origin. The Kirby-Bauer disc diffusion method was used to determine the antibiotic resistance profile of the isolates. Essential oil (EO) from the peels of lime purchased at a market in Ile-Ife was extracted by the hydro-distillation method, while the sensitivity of the isolates to EO was done via agar well diffusion method. The minimum inhibitory and minimum bactericidal concentrations (MIC and MBC) of the EO against the tested isolates were determined following standard methods. All the tested isolates exhibited multidrug resistance (MDR) characteristics. The multiple antibiotics resistant indexes (MARI%) for Gram-positive bacterial isolates ranged between 70% and 100% while that of Gram-negative was 100%. The yield of EO was 1% and the EO demonstrated activities at 25%, 50% and 100% v/v against the MDR bacterial isolates. The activity of EO was mostly not significantly different at the same concentration for all the isolates, and at different concentrations for each of the isolates. The MIC range for Gram-negative and Gram-positive isolates was between 0.195% to 3.125% v/v and 0.39% to 3.125% respectively while the range was between 1.563% to 3.125% and 0.781% to 6.250% v/v for MBC respectively. The study showed that EO from the peel of lime fruits demonstrated excellent in vitro antibacterial activity against MDR bacterial isolates. This potential can be further explored as an alternative for the treatment and management of infections caused by MDR bacterial isolates.