Insecticide resistance in Anopheles gambiae from the northern Democratic Republic of Congo, with extreme knockdown resistance (kdr) mutation frequencies revealed by a new diagnostic assay.
Insecticide resistance in Anopheles gambiae from the northern Democratic Republic of Congo, with extreme knockdown resistance (kdr) mutation frequencies revealed by a new diagnostic assay.
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DOI：
10.1186/s12936-018-2561-5
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发表时间：
2018-11-06
期刊：
影响因子：
3
通讯作者：
Weetman D
中科院分区：
文献类型：
--
作者：
Lynd A;Oruni A;Van't Hof AE;Morgan JC;Naego LB;Pipini D;O'Kines KA;Bobanga TL;Donnelly MJ;Weetman D
密码子1014电压门控钠通道的突变使多种昆虫对拟除虫菊酯产生敲除抗性（kdr）。冈比亚按蚊在密码子1014处表现出丝氨酸和苯丙氨酸两个突变等位基因；现在这两种疾病在非洲都很普遍。现有的筛选方法每次检测只能检测到一个耐药等位基因。建立了一种同时检测突变型等位基因和野生型等位基因的锁定核酸（LNA） qPCR方法。该三等位基因检测方法作为安家蚕杀虫剂抗性研究的一部分进行了评估。在刚果民主共和国北乌班吉以前未取样的地区发现严格的冈比亚菌（s.s.s）。使用世卫组织生物测定法对三个地点的样本进行了杀虫剂敏感性测试，在接触拟除虫菊酯之前使用增效剂PBO和不使用增效剂PBO，随后分析了Vgsc-1014和Vgsc-N1575Y突变的频率和耐药性相关性。将LNA-kdr 1014检测结果与标准TaqMan-kdr检测结果进行比较。到目前为止，捕获的主要媒介是冈比亚按蚊（84%），乌班吉北部仅检测到低频率的富氏按蚊（9%）。分子鉴定发现安。冈比亚疟蚊为主要媒介（99%），但科鲁兹按蚊的检出率很低。冈比亚按蚊对氨基甲酸酯类杀虫剂苯二威敏感，对滴滴涕和拟除虫菊酯、溴氰菊酯耐药。对这两种拟除虫菊酯的易感性在先前暴露于PBO后部分恢复，这表明可能与代谢抗性有关。冈比亚按蚊kdr耐药等位基因纯合子，同时存在L1014F和L1014S突变，N1575Y多态性出现频率较低。LNA-kdr试验同时检测到两种耐药等位基因，结果与两种TaqMan-kdr试验结果完全一致。该研究首次检测到N1575Y，为非洲中部按蚊的杀虫剂抗性和机制提供了罕见的数据。安省乌班吉北部人口。冈比亚虫表现出由代谢机制和Vgsc突变介导的杀虫剂抗性。LNA-kdr检测特别适用于1014S和1014F kdr等位基因共存的人群，并提供可靠的结果，具有更高的通量，成本仅为TaqMan检测的四分之一。本文的在线版本（10.1186/s12936-018-2561-5）包含补充材料，授权用户可使用。
Mutations in the voltage-gated sodium channel at codon 1014 confer knock-down resistance (kdr) to pyrethroids in a wide range of insects. Anopheles gambiae exhibits two mutant alleles at codon 1014, serine and phenylalanine; and both are now widespread across Africa. Existing screening methods only allow for one resistant allele to be detected per assay. A new locked nucleic acid (LNA) qPCR assay was developed for the simultaneous detection of both mutant alleles and the wild type allele in a single assay. This tri-allelic detection assay was assessed as part of a study of the insecticide resistance in An. gambiae sensu stricto (s.s.) in the previously un-sampled area of Nord Ubangi, Democratic Republic of the Congo. Samples from three sites were tested for insecticide susceptibility using WHO bioassays, with and without the synergist PBO preceding pyrethroid exposures, and were subsequently analysed for frequency and resistance-association of the Vgsc-1014 and Vgsc-N1575Y mutations. Results from the LNA-kdr 1014 assay were compared to results from standard TaqMan-kdr assays. Anopheles gambiae sensu lato (s.l.) was by far the predominant vector captured (84%), with only low frequencies of Anopheles funestus s.l. (9%) detected in Nord Ubangi. Molecular identification found An. gambiae s.s. to be the principal vector (99%) although Anopheles coluzzii was detected at very low frequency. Anopheles gambiae were susceptible to the carbamate insecticide bendiocarb, but resistant to DDT and to the pyrethroids permethrin and deltamethrin. Susceptibility to both pyrethroids was partially restored with prior exposure to PBO suggesting likely involvement of metabolic resistance. Anopheles gambiae s.s. was homozygous for kdr resistant alleles with both the L1014F and L1014S mutations present, and the N1575Y polymorphism was present at low frequency. The LNA-kdr assay simultaneously detected both resistant alleles and gave results entirely consistent with those from the two TaqMan-kdr assays. This study provides rare data on insecticide resistance and mechanisms in Anopheles from the centre of Africa, with the first detection of N1575Y. Nord Ubangi populations of An. gambiae s.s. show insecticide resistance mediated by both metabolic mechanisms and Vgsc mutations. The LNA-kdr assay is particularly suitable for use in populations in which both 1014S and 1014F kdr alleles co-occur and provides robust results, with higher throughput and at a quarter of the cost of TaqMan assays. The online version of this article (10.1186/s12936-018-2561-5) contains supplementary material, which is available to authorized users.
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影响因子：
3.2
作者：
Knox TB;Juma EO;Ochomo EO;Pates Jamet H;Ndungo L;Chege P;Bayoh NM;N'Guessan R;Christian RN;Hunt RH;Coetzee M
通讯作者：
Coetzee M
DOI：
10.1016/j.trstmh.2009.01.008
发表时间：
2009-11-01
影响因子：
2.2
作者：
Dabire, K. R.;Diabate, A.;Baldet, T.
通讯作者：
Baldet, T.
影响因子：
3
作者：
Bass, Chris;Nikou, Dimitra;Donnelly, Martin J.;Williamson, Martin S.;Ranson, Hilary;Ball, Amanda;Vontas, John;Field, Linda M.
通讯作者：
Field, Linda M.
影响因子：
64.8
作者：
Anopheles gambiae 1000 Genomes Consortium;Data analysis group;Partner working group;Sample collections—Angola:;Burkina Faso:;Cameroon:;Gabon:;Guinea:;Guinea-Bissau:;Kenya:;Uganda:;Crosses:;Sequencing and data production;Web application development;Project coordination
通讯作者：
Project coordination
影响因子：
3.7
作者：
Dabire, Roch K.;Namountougou, Moussa;Combary, Patrice
通讯作者：
Combary, Patrice