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Background: Knockdown resistance is conferred primarily by non-synonymous mutations that reduce pyrethroids binding to voltage-gated sodium channels. In 2014, kdr mutation V1016I in Aedes aegypti populations resistant to pyrethroids was reported for the first time in Colombiα, in 2016 another kdr mutation, F1534C, and in 2018 the mutation V419L. Nine populations of A. aegypti, previously characterized as being resistant to λ-cyhalothrin, deltamethrin, cyfluthrin and permethrin, were used for this study. Genomic DNA was used to determine genotypes by allele-specific PCR for mutations V1016I and F1534C, and to determine their association with pyrethroid resistance., Results: All the populations analyzed showed both mutations, with allelic frequencies of 0.07-0.35 for I1016 and 0.47-0.88 for C1534. A percentage of co-occurrence of mutant homozygotes I1016/C1534 of 5.3% was detected. A significant positive correlation was found between the frequency of the genotype I1016 and the resistance to permethrin, λ-cyhalothrin and cyfluthrin, but not to deltamethrin; on the other hand, the correlation was not significant for the C1534 genotype and the four pyrethroids evaluated. No significant correlation was found between the frequencies of the mutations V1016I and F1534C., Conclusions: Both mutations V1016I and F1534C are present in A. aegypti populations of the Colombian Caribbean, and although the frequency of F1534C exceeds V1016I, the latter was correlated to resistance to pyrethroid insecticides. © 2018 Society of Chemical Industry., (© 2018 Society of Chemical Industry.)