Identification and validation of internal reference genes for real-time quantitative polymerase chain reaction-based studies in Hyalomma anatolicum ticks

Cristiano Salata; Vanessa Monteil; Mikael Leijon; Lesley Bell-Sakyi; Ali Mirazimi

doi:10.1016/j.ttbdis.2020.101417

Identification and validation of internal reference genes for real-time quantitative polymerase chain reaction-based studies in Hyalomma anatolicum ticks

Ticks Tick Borne Dis. 2020 Jul;11(4):101417. doi: 10.1016/j.ttbdis.2020.101417. Epub 2020 Mar 19.

Authors

Cristiano Salata¹, Vanessa Monteil², Mikael Leijon³, Lesley Bell-Sakyi⁴, Ali Mirazimi⁵

Affiliations

¹ Department of Molecular Medicine, University of Padova, Via Gabelli, 63, IT-35121 Padova, Italy; Department of Microbiology, The Public Health Agency of Sweden, Nobels Väg 18, SE-171 82 Solna, Sweden. Electronic address: cristiano.salata@unipd.it.
² Department of Microbiology, The Public Health Agency of Sweden, Nobels Väg 18, SE-171 82 Solna, Sweden; Department for Laboratory Medicine, Karolinska University Hospital and KI, SE-14186 Huddinge, Sweden. Electronic address: vanessa.monteil@ki.se.
³ National Veterinary Institute, SE-756 51 Uppsala, Sweden. Electronic address: mikael.leijon@sva.se.
⁴ Department of Infection Biology, Institute of Infection and Global Health, University of Liverpool, Liverpool, Science Park IC2, 146 Brownlow Hill, Liverpool L3 5RF, United Kingdom. Electronic address: L.Bell-Sakyi@liverpool.ac.uk.
⁵ Department of Microbiology, The Public Health Agency of Sweden, Nobels Väg 18, SE-171 82 Solna, Sweden; Department for Laboratory Medicine, Karolinska University Hospital and KI, SE-14186 Huddinge, Sweden; National Veterinary Institute, SE-756 51 Uppsala, Sweden. Electronic address: Ali.Mirazimi@ki.se.

Abstract

Crimean-Congo hemorrhagic fever (CCHF) is an emerging tick-borne viral disease caused by the orthonairovirus CCHF virus (CCHFV). Ticks of the genus Hyalomma are the viral reservoir and they represent the main vector transmitting the virus to their hosts during blood feeding. However, how CCHFV replicates in its natural arthropod host cells and the nature of virus/host interactions are still largely unknown. With the aim of developing tools for use in this field, we identified and validated expression of four candidate endogenous control tick genes in a Hyalomma anatolicum-derived cell line. These genes will be useful for normalization of viral/cellular transcripts in infection/expression studies or as internal controls in molecular epidemiology surveys of pathogens transmitted by Hyalomma ticks.

Keywords: CCHFV; Endogenous genes; Hyalomma; Nairovirus; Real-Time RT-PCR; Reference genes; Tick cell line; Ticks.

Publication types

Research Support, Non-U.S. Gov't
Validation Study

MeSH terms

Animals
Cell Line / metabolism
Cell Line / virology
Hemorrhagic Fever Virus, Crimean-Congo / physiology*
Host Microbial Interactions
Ixodidae / genetics*
Real-Time Polymerase Chain Reaction / instrumentation
Real-Time Polymerase Chain Reaction / methods*
Real-Time Polymerase Chain Reaction / veterinary