The two most efficient and most recently radiated Afrotropical vectors of human malaria – Anopheles coluzzii and An. gambiae – are identified by single-locus diagnostic PCR assays based on species-specific markers in a 4 Mb region on chromosome-X centromere. Inherently, these diagnostic assays cannot detect interspecific autosomal admixture shown to be extensive at the westernmost and easternmost extremes of the species range. The main aim of this study was to develop novel, easy-to-implement tools for genotyping An. coluzzii and An. gambiae-specific ancestral informative markers (AIMs) identified from the Anopheles gambiae 1000 genomes (Ag1000G) project. First, we took advantage of this large set of data in order to develop a multilocus approach to genotype 26 AIMs on all chromosome arms valid across the species range. Second, we tested the multilocus assay on samples from Guinea Bissau, The Gambia and Senegal, three countries spanning the westernmost hybridization zone, where conventional species diagnostic is problematic due to the putative presence of a novel “hybrid form”. The multilocus assay was able to capture patterns of admixture reflecting those revealed by the whole set of AIMs and provided new original data on interspecific admixture in the region. Third, we developed an easy-to-use, cost-effective PCR approach for genotyping two AIMs on chromosome-3 among those included in the multilocus approach, opening the possibility for advanced identification of species and of admixed specimens during routine large scale entomological surveys, particularly, but not exclusively, at the extremes of the range, where WGS data highlighted unexpected autosomal admixture.

中文翻译：

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新的基因分型方法可轻松检测主要非洲热带疟疾载体物种，按蚊和 Anopheles coluzzii 之间的基因组混合物。冈比亚

人类疟疾的两种最有效和最近辐射的非洲热带病媒——Anopheles coluzzii和An。冈比亚——通过基于 X 染色体着丝粒上 4 Mb 区域中的物种特异性标记的单位点诊断 PCR 测定来鉴定。本质上，这些诊断测定不能检测在物种范围的最西端和最东端广泛存在的种间常染色体混合物。本研究的主要目的是开发用于基因分型An 的新颖、易于实施的工具。coluzzii和安。从冈比亚按蚊中鉴定出的冈比亚特异性祖先信息标记 (AIM)1000 个基因组（Ag1000G）项目。首先，我们利用这一庞大的数据集开发了一种多位点方法来处理跨物种范围有效的所有染色体臂上的基因型 26 AIM。其次，我们对来自几内亚比绍、冈比亚和塞内加尔的样本进行了多位点检测，这三个国家跨越最西部的杂交区，由于假定存在一种新的“杂交形式”，传统的物种诊断存在问题。多位点分析能够捕捉到混合物的模式，反映了整个 AIM 所揭示的模式，并提供了该地区种间混合物的新原始数据。第三，我们开发了一种易于使用、具有成本效益的 PCR 方法，用于对多位点方法中包含的 3 号染色体上的两个 AIM 进行基因分型，