Polymerase chain reaction (PCR)-based genotyping of mutations in the voltage-sensitive sodium channel (vssc) associated with resistance to pyrethroid insecticides is widely used and represents a potential early warning and monitoring system for insecticide resistance arising in mosquito populations, which are vectors of different human pathogens. In the secondary vector Aedes albopictus—an Asian species that has invaded and colonized the whole world, including temperate regions—sequencing of domain II of the vssc gene is still needed to detect the V1016G mutation associated with pyrethroid resistance. In this study we developed and tested a novel allele-specific PCR (AS-PCR) assay to genotype the V1016G mutation in this species and applied it to the analysis of wild populations from Italy. The results confirm the high accuracy of the novel AS-PCR and highlight frequencies of the V1016G allele as >5% in most sampling sites, with peaks of 20–45% in coastal touristic sites where pyrethroid treatments are extensively implemented, mostly for mosquito nuisance reduction. The high frequency of this mutation observed in Italian Ae. albopictus populations should serve as a warning bell, advocating for increased monitoring and management of a phenomenon which risks neutralizing the only weapon today available to counteract (risks of) arbovirus outbreaks.

中文翻译：

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一种新的等位基因特异性聚合酶链反应（AS-PCR）检测V1016G击倒抗性突变的方法证实了其在意大利伊蚊种群中的广泛存在

基于聚合酶链反应（PCR）的与对拟除虫菊酯类杀虫剂具有抗性相关的电压敏感钠通道（vssc）突变的基因分型已被广泛使用，它代表了潜在的预警和监测系统，该系统是蚊子种群中产生的杀虫剂抗性的媒介不同的人类病原体。在二级矢量纹伊蚊伊蚊已侵入和殖民统治整个世界-an亚洲种，包括域II的温带地区测序VSSC该基因仍需要检测与拟除虫菊酯抗性相关的V1016G突变。在这项研究中，我们开发并测试了一种新颖的等位基因特异性PCR（AS-PCR）分析，以对该物种中的V1016G突变进行基因分型，并将其应用于意大利野生种群的分析。结果证实了新型AS-PCR的高精度，并在大多数采样点中将V1016G等位基因的频率突出显示为> 5％，在广泛使用拟除虫菊酯治疗的沿海旅游点中，峰值为20–45％，主要用于蚊子滋扰减少。在意大利Ae中观察到此突变的频率很高。白化病 人口应作为警钟，主张加强对某种现象的监测和管理，这种现象可能会抵消当今唯一可用来抵制虫媒病毒爆发（的风险）的武器。