Knockdown resistance (kdr) gene of Aedes aegypti in Malaysia with the discovery of a novel regional specific point mutation A1007G.

BACKGROUND

Improved understanding of the molecular basis of insecticide resistance may yield new opportunities for control of relevant disease vectors. In this current study, we investigated the quantification responses for the phenotypic and genotypic resistance of Aedes aegypti populations from different states in Malaysia.

METHODS

We tested the insecticide susceptibility status of adult Ae. aegypti from populations of three states, Penang, Selangor and Kelantan (Peninsular Malaysia), against 0.25% permethrin and 0.25% pirimiphos-methyl using the World Health Organisation (WHO) adult bioassay method. Permethrin-resistant and -susceptible samples were then genotyped for domains II and III in the voltage-gated sodium channel (vgsc) gene using allele-specific polymerase chain reaction (AS-PCR) for the presence of any diagnostic single-nucleotide mutations. To validate AS-PCR results and to identify any possible additional point mutations, these two domains were sequenced.

RESULTS

The bioassays revealed that populations of Ae. aegypti from these three states were highly resistant towards 0.25% permethrin and 0.25% pirimiphos-methyl. Genotyping results showed that three knockdown (kdr) mutations (S989P, V1016G and F1534C) were associated with pyrethroid resistance within these populations. The presence of a novel mutation, the A1007G mutation, was also detected.

CONCLUSIONS

This study revealed the high resistance level of Malaysian populations of Ae. aegypti to currently used insecticides. The resistance could be due to the widespread presence of four kdr mutations in the field and this could potentially impact the vector control programmes in Malaysia and alternative solutions should be sought.