A next generation targeted amplicon sequencing method to screen for insecticide resistance mutations in Aedes aegypti populations reveals a rdl mutation in mosquitoes from Cabo Verde.
A next generation targeted amplicon sequencing method to screen for insecticide resistance mutations in Aedes aegypti populations reveals a rdl mutation in mosquitoes from Cabo Verde.
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DOI：
10.1371/journal.pntd.0010935
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发表时间：
2022-12
影响因子：
3.8
通讯作者：
Campino, Susana
中科院分区：
文献类型：
--
作者：
Collins, Emma L.;Phelan, Jody E.;Hubner, Magdalena;Spadar, Anton;Campos, Monica;Ward, Daniel;Acford-Palmer, Holly;Gomes, Ana Rita;Silva, Keily;Gomez, Lara Ferrero;Clark, Taane G.;Campino, Susana
伊蚊媒介传播许多全球健康问题的病毒，包括登革热、基孔肯雅热和寨卡病毒。这些媒介传播的病毒性疾病的治疗选择有限，疫苗的有效性也各不相同。因此，病媒综合管理是疾病控制的主要战略。然而，杀虫剂抗药性的日益出现和蔓延正在威胁病媒控制方法的效力。查明病媒种群中与抗药性有关的突变对于监测杀虫剂抗药性的发生和演变以及为控制战略提供信息十分重要。迫切需要快速和具有成本效益的基因组测序方法。在这里，我们提出了一种适应性强的靶向扩增子方法，用于在下一代测序平台中实现成本效益。这种方法可以识别单核苷酸多态性（SNP）和小插入和缺失（indels）的基因参与杀虫剂抗性在埃及伊蚊蚊子。我们设计并测试了11个扩增子，其包括ace-1（氨基甲酸酯靶标）、电压门控钠通道（vgsc;拟除虫菊酯、DDT和有机氯）和rdl（狄氏剂）基因的片段;从而覆盖了已建立的敲低抗性（kdr）突变（例如，S989 P、I1011 M/V、V1016 G/I和F1534 C），具有鉴定新的可能性。扩增子测定被设计有内部条形码，以促进以低成本多路复用大量蚊子，并使用Illumina平台测序。我们的方法在152 Ae上进行了评估。在卡波维德群岛收集的埃及蚊子，该群岛有虫媒病毒爆发的历史。扩增子序列数据揭示了146个SNPs，包括vgsc基因中的4个非同义多态性，ace-1和296 S rdl突变中的一个，这些突变先前与有机氯抗性相关。在98%的筛选蚊子中鉴定出296 S rdl突变，这与过去使用有机氯化合物（例如，滴滴涕）。总的来说，我们的工作表明，靶向扩增子测序是一种快速，稳健和具有成本效益的工具，可用于进行高通量监测杀虫剂抗性。许多病毒，如登革热和寨卡病毒，都是由埃及伊蚊传播的。这些病媒传播的疾病是全世界热带和亚热带地区的一个主要公共卫生问题。减轻其负担的主要战略是病媒控制，包括使用杀虫剂。然而，许多蚊子种群已经对杀虫剂产生了抗药性。在这里，我们提出了一种快速，强大，具有成本效益的工具，允许筛选与许多埃及伊蚊蚊子的杀虫剂抗性相关的基因，识别已知的抗性和新的突变。该检测将通过提供有关埃及伊蚊种群中杀虫剂抗性突变的出现和传播的信息，支持病媒控制策略。
Aedes mosquito vectors transmit many viruses of global health concern, including dengue, chikungunya and Zika. These vector-borne viral diseases have a limited number of treatment options, and vaccines vary in their effectiveness. Consequently, integrated vector management is a primary strategy for disease control. However, the increasing emergence and spread of insecticide resistance is threatening the efficacy of vector control methods. Identifying mutations associated with resistance in vector populations is important to monitor the occurrence and evolution of insecticide resistance and inform control strategies. Rapid and cost-effective genome sequencing approaches are urgently needed. Here we present an adaptable targeted amplicon approach for cost-effective implementation within next generation sequencing platforms. This approach can identify single nucleotide polymorphisms (SNPs) and small insertions and deletions (indels) in genes involved in insecticide resistance in Aedes aegypti mosquitoes. We designed and tested eleven amplicons, which included segments of the ace-1 (carbamate target), the Voltage-Gated Sodium Channel (vgsc; pyrethroids, DDT and organochlorines), and rdl (dieldrin) genes; thereby covering established knockdown resistance (kdr) mutations (e.g., S989P, I1011M/V, V1016G/I and F1534C), with the potential to identify novel ones. The amplicon assays were designed with internal barcodes, to facilitate multiplexing of large numbers of mosquitoes at low cost, and were sequenced using an Illumina platform. Our approach was evaluated on 152 Ae. aegypti mosquitoes collected in Cabo Verde, an archipelago with a history of arbovirus outbreaks. The amplicon sequence data revealed 146 SNPs, including four non-synonymous polymorphisms in the vgsc gene, one in ace-1 and the 296S rdl mutation previously associated with resistance to organochlorines. The 296S rdl mutation was identified in 98% of mosquitoes screened, consistent with the past use of an organochlorine compound (e.g., DDT). Overall, our work shows that targeted amplicon sequencing is a rapid, robust, and cost-effective tool that can be used to perform high throughput monitoring of insecticide resistance. Many viruses, such as dengue and Zika, are transmitted by Aedes aegypti mosquitoes. These vector-borne diseases are a major public health problem in the tropics and sub-tropics worldwide. The primary strategy to reduce their burden is vector control, including through the application of insecticides. However, many mosquito populations have developed resistance to insecticides. Here, we present a rapid, robust, and cost-effective tool that allows the screening of genes associated with insecticide resistance across many Aedes aegypti mosquitoes, identifying known resistance and novel mutations. This assay will support vector control strategies by informing on the emergence and spread of insecticide resistance mutations across Aedes aegypti populations.
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人类的影响塑造了埃及埃及，登革热和黄热病蚊子的历史和最新进化。
DOI：
10.1111/evo.12281
发表时间：
2014-02
期刊：
Evolution; international journal of organic evolution
影响因子：
--
作者：
Brown JE;Evans BR;Zheng W;Obas V;Barrera-Martinez L;Egizi A;Zhao H;Caccone A;Powell JR
通讯作者：
Powell JR
影响因子：
3
作者：
Granada Y;Mejía-Jaramillo AM;Strode C;Triana-Chavez O
通讯作者：
Triana-Chavez O
影响因子：
3.8
作者：
Achee NL;Gould F;Perkins TA;Reiner RC Jr;Morrison AC;Ritchie SA;Gubler DJ;Teyssou R;Scott TW
通讯作者：
Scott TW
影响因子：
1.9
作者：
Brengues, C;Hawkes, NJ;Hemingway, J
通讯作者：
Hemingway, J
影响因子：
14.9
作者：
Esling P;Lejzerowicz F;Pawlowski J
通讯作者：
Pawlowski J