Development of twenty-four microsatellite markers for Afrotropical Ornithodoros ticks

Background: Soft ticks of the genus Ornithodoros are responsible for the maintenance and transmission of the African swine fever (ASF) virus in the sylvatic and domestic viral cycles in Southern Africa. They are also the main vectors of Borrelia species causing relapsing fevers. Currently, no genetic markers are available for Afrotropical Ornithodoros ticks. As ASF spreads globally, such markers are needed to assess the role of ticks in the emergence of new outbreaks. The aim of this study was to design microsatellite markers that could be used for ticks of the Ornithodoros moubata complex, particularly Ornithodoros phacochoerus, to assess population structure and tick movements in ASF endemic areas. Methods: One hundred and fifty-one markers were designed using the O. moubata and O. porcinus genomes after elimination of repeated sequences in the genomes. All designed markers were tested on O. phacochoerus and O. porcinus DNA to select the best markers. Results: Twenty-four microsatelli..., This dataset results from genotyping of twenty-four microsatellite loci on Ornithodoros phacochoerus ticks and on four other Ornithodoros species. Fluorescent-labeled forward primers (FAM, VIC, NED or PET) were used for the 24 microsatellite loci. Touchdown PCRs were performed in six multiplexes of four microsatellite loci each. The amplification mix consisted in 2 μL of DNA template, 10 μL of 2x Type-it Microsatellite PCR Kit (Qiagen, Courtaboeuf, France), adjusted volume of fluorescence-labeled forward primer, and reverse primer for microsatellite loci, in a final volume of 20 μL. The touchdown PCR program was set as follows: 95°C for 3 min, then 10 cycles of 95°C for 20 sec, 60°C -0.5°C/cycle, for 30 sec, and 72°C for 1 min, then 30 cycles of 95°C for 20 sec, 55°C for 30 sec, and 72°C for 1 min, followed by a final extension step at 72°C for 7 min. Formamide for denaturation and GeneScan-600 (LIZ) Size Standard Kit for ladder were added to the PCR products before genotyping by capill..., , # Development of twenty-four microsatellite markers for Afrotropical Ornithodoros ticks ### Summary The three datasheets made available here result from the genotyping of twenty-four microsatellite loci on five *Ornithodoros* species.\ These datasheets can be transformed (using CREATE for instance) to be used in any population genetics software (Fstat, Genepop, Micro-Checker...). This dataset was used to test the robustness of twenty-four microsatellite loci for the species *Ornithodoros phacochoerus*. The microsatellite loci were also tested on four other *Ornithodoros* species (*O. moubata*, *O. porcinus*, *O. maritimus*, and *O. erraticus*) to assess specificity. ## Description of the data and file structure This dataset contains three different datasheets of genotyping results. Two of the datasheets present genotyping data from the species *O. phacochoerus*. *Ornithodoros phacochoerus* ticks came from two different sites: Coutada9 Game Reserve (pop 69) and Gorongosa National Pa...# 非洲热带区钝缘蜱24个微卫星标记的开发 ## 背景 钝缘蜱属（Ornithodoros）软蜱是非洲南部丛林与家养病毒循环中非洲猪瘟（African swine fever, ASF）病毒维持与传播的关键媒介，同时也是引发回归热的疏螺旋体属（Borrelia）物种的主要传播媒介。目前，非洲热带区钝缘蜱尚无可用的遗传标记。随着非洲猪瘟在全球范围内扩散，亟需此类标记以评估蜱虫在新发疫情中的作用。本研究旨在为穆巴钝缘蜱复合组（Ornithodoros moubata complex）的蜱虫，尤其是猪钝缘蜱（Ornithodoros phacochoerus），设计可用于评估非洲猪瘟流行区蜱种群结构与移动模式的微卫星标记。 ## 方法 本研究基于穆巴钝缘蜱（O. moubata）与豚钝缘蜱（O. porcinus）的基因组序列，在剔除基因组内重复序列后共设计了151个分子标记。所有设计得到的标记均在猪钝缘蜱（O. phacochoerus）与豚钝缘蜱（O. porcinus）的DNA样本中进行扩增验证，以筛选出最优标记。 ## 结果 本数据集基于24个微卫星位点，对猪钝缘蜱（Ornithodoros phacochoerus）及另外4种钝缘蜱进行基因分型得到。 24个微卫星位点均采用荧光标记正向引物（FAM、VIC、NED或PET）进行扩增。实验采用6组多重PCR体系，每组包含4个微卫星位点。 扩增反应体系总体积为20 μL，包含2 μL DNA模板、10 μL 2×Type-it微卫星PCR试剂盒（Qiagen，法国库塔伯夫）、调整体积的荧光标记正向引物与微卫星位点反向引物。降落PCR程序设置如下：95℃预变性3 min；随后进行10个循环，每个循环为95℃变性20 s、60℃退火30 s（每个循环退火温度降低0.5℃）、72℃延伸1 min；再进行30个循环，每个循环为95℃变性20 s、55℃退火30 s、72℃延伸1 min；最后72℃终延伸7 min。 基因分型前，向PCR产物中加入变性用甲酰胺与分子量内标GeneScan-600（LIZ）分子量标准试剂盒，随后通过毛细管电泳完成基因分型。 ### 概述 本数据集提供的3份数据文件均基于5种钝缘蜱（Ornithodoros）的24个微卫星位点基因分型结果生成。 这些数据文件可通过格式转换（例如使用CREATE工具）适配于任意种群遗传学分析软件（如Fstat、Genepop、Micro-Checker等）。 本数据集用于验证24个微卫星位点在猪钝缘蜱（Ornithodoros phacochoerus）中的扩增稳定性，同时还在另外4种钝缘蜱（穆巴钝缘蜱O. moubata、豚钝缘蜱O. porcinus、滨海钝缘蜱O. maritimus与奇异钝缘蜱O. erraticus）中开展了特异性验证。 ## 数据与文件结构说明 本数据集包含3份不同的基因分型结果数据文件，其中2份来自猪钝缘蜱（O. phacochoerus）的基因分型数据。该物种的蜱虫样本采集自两个不同采样点：Coutada9禁猎区（种群69）与戈龙戈萨国家公园（Gorongosa National Pa...）。