# Development and Characterization of Microsatellite Genetic Markers for Hyalomma rufipes, a Tick Vector of Crimean‐Congo Hemorrhagic Fever Virus

## TL;DR

Researchers created 14 genetic markers to study the population structure of a tick species that spreads a dangerous virus in East Africa.

## Contribution

Development of 14 polymorphic microsatellite markers for population genetic studies of Hyalomma rufipes.

## Key findings

- 14 microsatellite markers were developed and validated for H. rufipes with high allelic richness and gene diversity.
- Markers showed signs of null alleles but no stuttering or allelic dropout, making them suitable for population studies.
- These markers will help understand the dispersal and genetic structure of H. rufipes in relation to CCHFV transmission.

## Abstract

Hyalomma rufipes
is a widely distributed tick species and a competent vector of Crimean‐Congo Hemorrhagic Fever Virus (CCHFV), a serious zoonotic pathogen endemic to over 30 countries. Despite the epidemiological importance of CCHFV and
H. rufipes
in East Africa, little is known about the genetic structure and movement of
H. rufipes
populations, limiting the understanding of CCHFV transmission dynamics in this region. This study developed and characterized 14 polymorphic microsatellite markers to support population genetic studies of
H. rufipes
.
H. rufipes
ticks were collected from livestock in Garissa and Isiolo counties in northern Kenya. Morphological identification was confirmed using 16S rRNA Sanger sequencing and phylogenetic analysis. Low‐pass whole genome sequencing was performed on representative samples, and the Quality and Diversity of DNA (QDD) pipeline was used to identify and design microsatellite primers. Of 59,201 candidate loci, 30 were selected for initial screening; 14 loci consistently amplified and were polymorphic. These included mostly tetranucleotide repeats and showed high allelic richness and gene diversity. Several loci showed signs of null alleles, but no evidence of stuttering or allelic dropout was found. These newly developed microsatellite markers provide a valuable tool for investigating
H. rufipes
population dynamics and dispersal, with the ultimate goal of understanding CCHFV transmission dynamics in East Africa.

This study developed and validated 14 polymorphic microsatellite markers to investigate the population genetics of Hyalomma rufipes, a tick species that transmits Crimean‐Congo Hemorrhagic Fever Virus (CCHFV) in East Africa. Ticks collected from livestock in northern Kenya were morphologically and genetically identified, followed by genome sequencing and microsatellite marker design. The markers demonstrated high genetic diversity and will aid in understanding the movement and population structure of H. rufipes, which is crucial for mapping CCHFV transmission in the region.