This study was conducted to evaluate the phytochemical composition and larvicidal effect of leaf essential oil from Murraya exotica against early fourth-instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. Gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS) analyses revealed that the essential oil contained 27 components. The major chemical components identified were β-humulene (40.62%), benzyl benzoate (23.96%), β-caryophyllene (7.05%) and α-terpinene (5.66%). The larval mortality was observed after 12 and 24 h of exposure period. The results revealed that essential oil showed varied levels of larvicidal activity against A. aegypti, A. stephensi and C. quinquefasciatus. After 12 h of exposure period, the larvicidal activities were LC₅₀ = 74.7 and LC₉₀ = 152.7 ppm (A. aegypti), LC₅₀ = 56.3 and LC₉₀ = 107.8 ppm (A. stephensi ), and LC₅₀ = 74.4 and LC₉₀ = 136.9 ppm (C. quinquefasciatus) and the larvicidal activities after 24 h of exposure period were LC₅₀ = 35.8 and LC₉₀ = 85.4 ppm (A. aegypti), LC₅₀ = 31.3 and LC₉₀ = 75.1 ppm (A. stephensi), and LC₅₀ = 43.2 and LC₉₀ = 103.2 ppm (C. quinquefasciatus). These results suggest that leaf essential oil from M. exotica is a promising and eco-friendly source of natural larvicidal agent against A. aegypti, A. stephensi and C. quinquefasciatus.