Harnessing Attenuation-Related Mutations of Viral Genomes: Development of a Serological Assay to Differentiate between Capripoxvirus-Infected and -Vaccinated Animals
Francisco J. Berguido,
Tesfaye Rufael Chibssa,
Angelika Loitsch,
Yang Liu,
Kiril Krstevski,
Igor Djadjovski,
Eeva Tuppurainen,
Tamaš Petrović,
Dejan Vidanović,
Philippe Caufour,
Tirumala Bharani K. Settypalli,
Clemens Grünwald-Gruber,
Reingard Grabherr,
Adama Diallo,
Giovanni Cattoli,
Charles Euloge Lamien
Affiliations
Francisco J. Berguido
Animal Production and Health Laboratory, Animal Production and Health Section, Joint FAO/IAEA Division, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, WagramerStrasse 5, P.O. Box 100, 1400 Vienna, Austria
Tesfaye Rufael Chibssa
Animal Health Institute, Sebeta P.O. Box 4, Ethiopia
Angelika Loitsch
Austrian Agency for Health and Food Safety (AGES), Spargelfeldstrasse 191, 1220 Vienna, Austria
Yang Liu
China National Clinical Research Center for Neurological Diseases, Beijing Tiantan Hospital, Capital Medical University, Beijing 100070, China
Kiril Krstevski
Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, 1000 Skopje, North Macedonia
Igor Djadjovski
Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, 1000 Skopje, North Macedonia
Eeva Tuppurainen
Institute of International Animal Health/One Health, Friedrich-Loeffler-Institut, 17493 Greifswald, Germany
Tamaš Petrović
Scientific Veterinary Institute “Novi Sad”, 21000 Novi Sad, Serbia
Dejan Vidanović
Veterinary Specialized Institute Kraljevo, Zicka 34, 36103 Kraljevo, Serbia
Philippe Caufour
UMR ASTRE Cirad-Inrae, University of Montpellier (I-MUSE), 34398 Montpellier, France
Tirumala Bharani K. Settypalli
Animal Production and Health Laboratory, Animal Production and Health Section, Joint FAO/IAEA Division, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, WagramerStrasse 5, P.O. Box 100, 1400 Vienna, Austria
Clemens Grünwald-Gruber
Core Facility Mass Spectrometry, University of Natural Resources and Life Sciences Vienna, 1190 Vienna, Austria
Reingard Grabherr
Institute of Biotechnology, University of Natural Resources and Life Sciences (BOKU), Muthgasse 18, 1190 Vienna, Austria
Adama Diallo
Independent Researcher, Hahngasse, 24-26, 02/07, 1090 Vienna, Austria
Giovanni Cattoli
Animal Production and Health Laboratory, Animal Production and Health Section, Joint FAO/IAEA Division, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, WagramerStrasse 5, P.O. Box 100, 1400 Vienna, Austria
Charles Euloge Lamien
Animal Production and Health Laboratory, Animal Production and Health Section, Joint FAO/IAEA Division, Department of Nuclear Sciences and Applications, International Atomic Energy Agency, WagramerStrasse 5, P.O. Box 100, 1400 Vienna, Austria
Sheeppox, goatpox, and lumpy skin disease caused by the sheeppox virus (SPPV), goatpox virus (GTPV), and lumpy skin disease virus (LSDV), respectively, are diseases that affect millions of ruminants and many low-income households in endemic countries, leading to great economic losses for the ruminant industry. The three viruses are members of the Capripoxvirus genus of the Poxviridae family. Live attenuated vaccines remain the only efficient means for controlling capripox diseases. However, serological tools have not been available to differentiate infected from vaccinated animals (DIVA), though crucial for proper disease surveillance, control, and eradication efforts. We analysed the sequences of variola virus B22R homologue gene for SPPV, GTPV, and LSDV and observed significant differences between field and vaccine strains in all three capripoxvirus species, resulting in the truncation and absence of the B22R protein in major vaccines within each of the viral species. We selected and expressed a protein fragment present in wildtype viruses but absent in selected vaccine strains of all three species, taking advantage of these alterations in the B22R gene. An indirect ELISA (iELISA) developed using this protein fragment was evaluated on well-characterized sera from vaccinated, naturally and experimentally infected, and negative cattle and sheep. The developed wildtype-specific capripox DIVA iELISA showed >99% sensitivity and specificity for serum collected from animals infected with the wildtype virus. To the best of our knowledge, this is the first wildtype-specific, DIVA-capable iELISA for poxvirus diseases exploiting changes in nucleotide sequence alterations in vaccine strains.
