High-throughput barcoding method for the genetic surveillance of insecticide resistance and species identification in Anopheles gambiae complex malaria vectors.
High-throughput barcoding method for the genetic surveillance of insecticide resistance and species identification in Anopheles gambiae complex malaria vectors.
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DOI：
10.1038/s41598-022-17822-8
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发表时间：
2022-08-16
影响因子：
4.6
通讯作者：
Campino, Susana
中科院分区：
文献类型：
--
作者：
Campos, Monica;Phelan, Jody;Spadar, Anton;Collins, Emma;Goncalves, Aderitow;Pelloquin, Bethanie;Vaselli, Natasha Marcella;Meiwald, Anne;Clark, Emma;Stica, Caleb;Orsborne, James;Sylla, Moussa;Edi, Constant;Camara, Denka;Mohammed, Abdul Rahim;Afrane, Yaw Asare;Kristan, Mojca;Walker, Thomas;Gomez, Lara Ferrero;Messenger, Louisa A.;Clark, Taane G.;Campino, Susana
监测疟疾病媒种类和监测杀虫剂抗药性对于制定疟疾控制战略和支持减少感染和疾病至关重要。蚊子的遗传条形码是帮助高通量监测杀虫剂耐药性、区分兄弟物种和检测是否存在疟原虫感染的有用工具。在这项研究中，我们结合了多重PCR、定制设计的双重索引和Illumina下一代测序技术，对来自按蚊的四个物种进行了高通量单核苷酸多态(SNP)分析。冈比亚复合体(An.冈比亚感觉严格，一种。Coluzzii、An.Arabiens和An.Melas)。通过仅扩增和测序14个遗传片段(每个500个碱基)，我们能够同时检测到疟原虫感染；ACE1、GSTe2、vgsc和Rd1基因中与杀虫剂抗性相关的SNPs；核糖体内部转录间隔区(ITS1和ITS2)和基因间隔区(IGS)的部分序列；短散布元件(SINE)；以及用于物种鉴定和遗传多样性的线粒体基因(Cox1和ND4)。使用该扩增序列方法对四个选定的AN进行测序。针对冈比亚复杂种，我们在杀虫剂靶标基因中发现了15个非同义突变，其中包括与抗性相关的突变和两个新的突变(vgsc中的F1525L和GSTe2中的D148E)。总体而言，我们提供了一个可靠且具有成本效益的高通量面板来监控AN。疟疾流行地区冈比亚亚纲复合蚊子。
Surveillance of malaria vector species and the monitoring of insecticide resistance are essential to inform malaria control strategies and support the reduction of infections and disease. Genetic barcoding of mosquitoes is a useful tool to assist the high-throughput surveillance of insecticide resistance, discriminate between sibling species and to detect the presence of Plasmodium infections. In this study, we combined multiplex PCR, custom designed dual indexing, and Illumina next generation sequencing for high throughput single nucleotide polymorphism (SNP)-profiling of four species from the Anopheles (An.) gambiae complex (An. gambiae sensu stricto, An. coluzzii, An. arabiensis and An. melas). By amplifying and sequencing only 14 genetic fragments (500 bp each), we were able to simultaneously detect Plasmodium infection; insecticide resistance-conferring SNPs in ace1, gste2, vgsc and rdl genes; the partial sequences of nuclear ribosomal internal transcribed spacers (ITS1 and ITS2) and intergenic spacers (IGS), Short INterspersed Elements (SINE), as well as mitochondrial genes (cox1 and nd4) for species identification and genetic diversity. Using this amplicon sequencing approach with the four selected An. gambiae complex species, we identified a total of 15 non-synonymous mutations in the insecticide target genes, including previously described mutations associated with resistance and two new mutations (F1525L in vgsc and D148E in gste2). Overall, we present a reliable and cost-effective high-throughput panel for surveillance of An. gambiae complex mosquitoes in malaria endemic regions.
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影响因子：
3.7
作者：
Boissière A;Gimonneau G;Tchioffo MT;Abate L;Bayibeki A;Awono-Ambéné PH;Nsango SE;Morlais I
通讯作者：
Morlais I
非洲瞬间蚊子（双翅目：Culicidae）对疟疾控制程序的形态鉴定的重要性。
影响因子：
3
作者：
Erlank E;Koekemoer LL;Coetzee M
通讯作者：
Coetzee M
影响因子：
10.7
作者：
Grau-Bové X;Tomlinson S;O'Reilly AO;Harding NJ;Miles A;Kwiatkowski D;Donnelly MJ;Weetman D;Anopheles gambiae 1000 Genomes Consortium
通讯作者：
Anopheles gambiae 1000 Genomes Consortium
影响因子：
3
作者：
da Cruz, Derciliano Lopes;Santos Paiva, Marcelo Henrique;Junqueira Ayres, Constancia Flavia
通讯作者：
Junqueira Ayres, Constancia Flavia
影响因子：
14.9
作者：
Esling P;Lejzerowicz F;Pawlowski J
通讯作者：
Pawlowski J