Rhipicephalus sanguineus amplicon sequencing
Multiplexed amplicon short-read sequences of four phylogenetically informative mitochondrial (COI, 12S, and 16S) and nuclear (ITS2) loci, and five loci targeting regions encompassing common genetic mutations in the voltage-gated sodium channel (VGSC) and GABA-gated chloride channel (GABA-Cl) genes associated with synthetic pyrethroid (SP) and dieldrin/fipronil acaricide resistance. Two separate primer pairs capture the region of the domain III segment 6 of the VGSC gene containing the single point mutation (T2134C) previously associated with resistance to SPs in Rh. sanguineus s.l. from the US. Also included is a 199bp region of domain II segment 6 of the VGSC gene encompassing the single SNP associated with the SP resistance substitution L1014F, and a 167bp region of domain II segment 4-5 that encompasses regions containing eight other known mutations associated with SP resistance in a variety of arthropod species, including Rh. sanguineus s.l. Also included is a 231bp region of the GABA-Cl gene that contains several mutations associated with dieldrin/fipronil resistance in Drosophila spp. and Rh. microplus.