A targeted amplicon sequencing panel to simultaneously identify mosquito species and Plasmodium presence across the entire Anopheles genus.
A targeted amplicon sequencing panel to simultaneously identify mosquito species and Plasmodium presence across the entire Anopheles genus.
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DOI：
10.1111/1755-0998.13436
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发表时间：
2022-01
影响因子：
7.7
通讯作者：
Lawniczak MKN
中科院分区：
文献类型：
--
作者：
Makunin A;Korlević P;Park N;Goodwin S;Waterhouse RM;von Wyschetzki K;Jacob CG;Davies R;Kwiatkowski D;St Laurent B;Ayala D;Lawniczak MKN
按蚊是一种多样的蚊子属，包括超过500种描述的物种，包括所有已知的人类疟疾媒介。虽然对数量有限的关键病媒物种进行了详细研究，但消灭疟疾的目标要求对所有潜在病媒物种进行监测。在这里，我们开发了一种多位点扩增子测序方法，该方法针对按蚊基因组中的62个高度可变的位点和疟原虫基因组中的两个保守位点，同时揭示了蚊子的种类以及蚊子是否携带疟疾寄生虫。我们还开发了一种廉价，非破坏性和高通量的DNA提取工作流程，为多重PCR提供来自单个蚊子的模板DNA，这意味着可以返回产生意外结果的标本进行形态学检查。在一次MiSeq运行中可以对1000多只蚊子进行测序，我们展示了该小组使用来自非洲，东南亚和南美洲的40种物种的测序数据分配物种身份的能力。我们还表明，该方法可以用来解决地理种群结构内的一个。冈比亚和安. coluzzii种群，因为基于来自1000多只蚊子的这62个基因座确定的种群结构密切反映了通过全基因组测序揭示的情况。这种端到端的方法快速、廉价、可靠且准确，这使其成为非常有前途的大规模蚊子遗传监测和病媒控制技术。
Anopheles is a diverse genus of mosquitoes comprising over 500 described species, including all known human malaria vectors. While a limited number of key vector species have been studied in detail, the goal of malaria elimination calls for surveillance of all potential vector species. Here, we develop a multilocus amplicon sequencing approach that targets 62 highly variable loci in the Anopheles genome and two conserved loci in the Plasmodium mitochondrion, simultaneously revealing both the mosquito species and whether that mosquito carries malaria parasites. We also develop a cheap, nondestructive, and high-throughput DNA extraction workflow that provides template DNA from single mosquitoes for the multiplex PCR, which means specimens producing unexpected results can be returned to for morphological examination. Over 1000 individual mosquitoes can be sequenced in a single MiSeq run, and we demonstrate the panel’s power to assign species identity using sequencing data for 40 species from Africa, Southeast Asia, and South America. We also show that the approach can be used to resolve geographic population structure within An. gambiae and An. coluzzii populations, as the population structure determined based on these 62 loci from over 1000 mosquitoes closely mirrors that revealed through whole genome sequencing. The end-to-end approach is quick, inexpensive, robust, and accurate, which makes it a promising technique for very large-scale mosquito genetic surveillance and vector control.
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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：
48
作者：
Callahan BJ;McMurdie PJ;Rosen MJ;Han AW;Johnson AJ;Holmes SP
通讯作者：
Holmes SP
影响因子：
4
作者：
Bell, AS;Ranford-Cartwright, LC
通讯作者：
Ranford-Cartwright, LC
DOI：
10.1093/bioinformatics/bts565
发表时间：
2012-12-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Fu L;Niu B;Zhu Z;Wu S;Li W
通讯作者：
Li W
DOI：
10.4269/ajtmh.2003.69.200
发表时间：
2003-08-01
影响因子：
3.3
作者：
Cohuet, A;Simard, F;Fontenille, D
通讯作者：
Fontenille, D