Development of twenty-four microsatellite markers for Afrotropical Ornithodoros ticks
Authors/Creators
- 1. Centre de Coopération Internationale en Recherche Agronomique pour le Développement
- 2. Agricultural Research Institute of Mozambique, Chimoio*
Description
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 microsatellite markers were genotyped on two populations of O. phacochoerus and on few individuals from four other Ornithodoros species. Nineteen markers were selected to be as robust as possible for population genetic studies on O. phacochoerus.
Conclusions: The microsatellite markers developed here represent the first genetic tool to study nidicolous populations of Afrotropical Ornithodoros. This dataset contains the genotyping results obtained for all twenty-four markers tested.
Notes
Funding provided by: National Institute of Food and Agriculture
Crossref Funder Registry ID: https://ror.org/05qx3fv49
Award Number:
Methods
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 capillary electrophoresis at the GPTR laboratory (Great Regional Technical Platform of genotyping, AGAP Institut/CIRAD, Montpellier, France) with an ABI 3500xL Genetic Analyzer (Applied Biosystems, Foster City, CA, USA).
Genotypes were read using GeneMapper® v.6 software (Applied Biosystems, Waltham, MA, USA). Allele bins were set manually after a review of all samples. Allele scoring was performed automatically according to the bin set designed for the marker, then manually checked by two different experimenters. Alleles were named according to their length in base pairs.
Files
Genotyping.zip
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