Crimean-Congo hemorrhagic fever (CCHF) is an emerging tick-borne zoonotic disease caused by the CCHF virus (CCHFV). In this study, an experimental approach combining RT-PCR and proteomics was used for the identification and characterization of CCHFV in 106 ticks from 7 species that were collected from small ruminants in Greece. The methodological approach included an initial screening for CCHFV by RT-PCR followed by proteomics analysis of positive and control negative tick samples. This novel approach allowed the identification of CCHFV-positive ticks and provided additional information to corroborate the RT-PCR findings using a different approach. Two ticks, Dermacentor marginatus and Haemaphysalis parva collected from a goat and a sheep, respectively were positive for CCHFV. The sequences for CCHFV RNA segments S and L were characterized by RT-PCR and proteomics analysis of tick samples, respectively. These results showed the possibility of combining analyses at the RNA and protein levels using RT-PCR and proteomics for the characterization of CCHFV in ticks. The results supported that the CCHFV identified in ticks are genetic variants of the AP92 strain. Although the AP92-like strains probably do not represent a high risk of CCHF to the population, the circulation of genetically diverse CCHFV strains could potentially result in the appearance of novel viral genotypes with increased pathogenicity and fitness.

Department of Microbiology Medical School Aristotle University of Thessaloniki 54124 Thessaloniki Greece

Institute of Parasitology Biology Center of the Academy of Sciences of the Czech Republic 37005 České Budějovice Czech Republic Faculty of Science University of South Bohemia České Budějovice Czech Republic

SaBio Instituto de Investigación de Recursos Cinegéticos IREC CSIC UCLM JCCM 13005 Ciudad Real Spain

SaBio Instituto de Investigación de Recursos Cinegéticos IREC CSIC UCLM JCCM 13005 Ciudad Real Spain Department of Veterinary Pathobiology Center for Veterinary Health Sciences Oklahoma State University Stillwater Oklahoma USA

Veterinary Research Institute Hellenic Agricultural Organisation Demeter NAGREF Campus 57001 Thermi Thessaloniki Greece

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