<p><i>Aedes aegypti</i>, the primary vector of dengue, has developed widespread resistance to insecticides, posing a threat to the efficacy of vector control programs. This study assessed the susceptibility status of <i>Ae. aegypti</i> across Malaysian dengue hotspots and characterized knockdown resistance (<i>kdr</i>) mutations, including the first detection of T1520I. Adult female mosquitoes were collected from seven localities across five Malaysian states. Susceptibility to deltamethrin (0.03%), permethrin (0.4%), pirimiphos-methyl (60&#xa0;mg/m²), and malathion (5%) was evaluated using bioassay protocols. Knockdown times (KT₅₀, KT₉₅) and resistance ratios (RR) were estimated using probit analysis, and the voltage-gated sodium channel gene was sequenced to detect <i>kdr</i> mutations. All field populations remained susceptible to malathion, with mortality ≥ 95%, though one Johor strain (ABS) showed suspected resistance (95%). Pirimiphos-methyl resistance was widespread, with mortality as low as 6% (ABS). Whereas resistance to pyrethroids was pronounced, with deltamethrin mortality ranging from 22% (ABS) to 95% (AA), and permethrin from 0% (TMB, ABS) to 95% (AA). Knockdown assays revealed high resistance ratios, particularly for permethrin (TMB: RR₅₀=9.98, RR₉₅=14.98; ABS: RR₉₅=41.46). Sequencing identified multiple <i>kdr</i> mutations, including S989P, V1016G, F1534C, A1007G, and the novel detection of T1520I in Malaysian populations. F1534C was strongly associated with permethrin resistance, while V1016G and S989P predominated under deltamethrin exposure. <i>Aedes aegypti</i> populations in Malaysia exhibit widespread pyrethroid resistance and emerging organophosphate resistance, underpinned by multiple <i>kdr</i> mutations. The first detection of T1520I underscores the evolving genetic basis of resistance. These findings highlight the urgent need for integrated resistance management, including molecular surveillance and insecticide rotation, to sustain effective dengue control.</p>