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Background  Mutation of the voltage-gated sodium channel (VGSC) gene, or knockdown resistance ( kdr ) gene, is an important resistance mechanism against DDT and pyrethroids for dengue vector  Aedes albopictus . A phenylalanine to serine (F1534S), leucine (F1534L) and cysteine (F1534C) substitution were detected in many  Ae. albopictus  populations around the world, and the mutant allele frequencies have been increasing in recent years. Therefore, it is essential to establish a simple, time-saving and cost-effective procedure to monitor the alleles in large-scale studies.    Methods  Based on the mutation genotypes of the 1534 locus in the  kdr  gene, F/F, F/S, F/C, F/L, S/S, C/C, L/L and S/C, we designed specific forward and reverse primers and optimized the reaction conditions for establishing of the allele-specific PCR(AS-PCR) detection technique. DNA sequencing in this study was taken as the gold standard, and used to determine the accuracy of AS-PCR.    Results  The designed AS-PCR technique showed high specificity for distinguishing the mutations at the 1534 locus, as the accuracy for F/F, F/S, F/C, F/L, S/S, C/C and S/C were 100%, 95.35%, 100%, 100%, 100%, 100% and 100%, respectively.    Conclusions  The designed AS-PCR technique effectively distinguished individual genotypes for the mutations at the 1534 locus in the  kdr  gene, which could facilitate the knockdown resistance surveillance in  Ae. albopictus  in large-scale studies  .

parasites and vectors

Establishment of an innovative and sustainable PCR technique for 1534 locus mutation of the knockdown resistance (kdr) gene in the dengue vector Aedes albopictus