Abstract
BackgroundThe spread of insecticide resistance (IR) is a major threat to vector control programmes for mosquito-borne diseases. Early detection of IR using diagnostic markers could help inform these programmes, especially in remote locations where gathering reliable bioassay data is challenging. Most current molecular tests for genetic IR markers are only suitable for use in well-equipped laboratory settings. There is an unmet need for field-applicable diagnostics.MethodsA single-cartridge test was designed to detect key IR mutations in the major African vector of malaria, Anopheles gambiae. Developed on the portable, rapid, point-of-care compatible PCR platform - Genedrive® (genedrive® plc), the test comprises two assays which target single nucleotide polymorphisms (SNPs) in the voltage gated sodium channel (VGSC) gene that exert interactive effects on knockdown resistance (kdr): L1014F, L1014S and N1575Y.ResultsDistinct melt peaks were observed for each allele at each locus. Preliminary validation of these assays using a test panel of 70 An. gambiae samples showed complete agreement of our assays with the widely-used TaqMan assays, achieving a sensitivity and specificity of 100%.ConclusionHere we show the development of an insecticide resistance detection assay for use on the Genedrive® platform that has the potential to be the first field-applicable diagnostic for kdr.
Highlights
The spread of insecticide resistance (IR) is a major threat to vector control programmes for mosquitoborne diseases
All Insecticide-treated nets (ITN) and Long-lasting insecticide treated nets (LLIN) were formulated with pyrethroid insecticides [2]
Primer and probe design Two separate assays were designed to target three individual Single nucleotide polymorphism (SNP) within the voltage-gated sodium channel (Vgsc) gene, the first to discriminate between the two mutations at the L1014 locus and the other at the 1575Y locus
Summary
The spread of insecticide resistance (IR) is a major threat to vector control programmes for mosquitoborne diseases. Control programmes for vector-borne diseases, such as malaria, are heavily reliant on the use of insecticides to reduce vector populations. The use of insecticide-treated nets (ITNs), long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) plays a major role in the overall reduction in global malaria burden. F and S mutations have both been detected across the continent and sometimes co-occur [8]. The mutations do not occur on the same haplotypes, but where both alleles are present, effects on kdr are additive [7, 9]. The 1575Y (‘Y’) mutation is only present on the F haplotype and acts as an amplifier of resistance to both pyrethroids and DDT [10, 11]
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