Multiplex Assay for Simultaneous Detection of Antibodies against Crimean-Congo Hemorrhagic Fever Virus Nucleocapsid Protein and Glycoproteins in Ruminants.
Hoste, Alexis; Djadjovski, Igor; Jiménez-Clavero, Miguel Ángelet al.
[en] Crimean-Congo hemorrhagic fever virus (CCHFV) is a widespread tick-borne zoonotic virus that causes Crimean-Congo hemorrhagic fever (CCHF). CCHF is asymptomatic in infected animals but can develop into severe illness in humans, with high case-fatality rates. Due to complex environmental and socio-economic factors, the distribution of CCHFV vectors is changing, leading to disease occurrence in previously unaffected countries. Neither an effective treatment nor a vaccine has been developed against CCHFV; thus, surveillance programs are essential to limit and control the spread of the virus. Furthermore, the WHO highlighted the need of assays that can cover a range of CCHFV antigenic targets, DIVA (differentiating infected from vaccinated animals) assays, or assays for future vaccine evaluation. Here, we developed a multiplex assay, based on a suspension microarray, able to detect specific antibodies in ruminants to three recombinantly produced CCHFV proteins: the nucleocapsid (N) protein and two glycoproteins, GN ectodomain (GNe), and GP38. This triplex assay was used to assess the antibody response in naturally infected animals. Out of the 29 positive field sera to the N protein, 40% showed antibodies against GNe or GP38, with 11 out of these 12 samples being positive to both glycoproteins. To determine the diagnostic specificity of the test, a total of 147 sera from Spanish farms free of CCHFV were included in the study. This multiplex assay could be useful to detect antibodies to different proteins of CCHFV as vaccine target candidates and to study the immune response to CCHFV in infected animals and for surveillance programs to prevent the further spread of the virus. IMPORTANCE Crimean-Congo hemorrhagic fever virus (CCHFV) causes Crimean-Congo hemorrhagic fever, which is one of the most important tick-borne viral diseases of humans and has recently been found in previously unaffected countries such as Spain. The disease is asymptomatic in infected animals but can develop into severe illness in humans. As neither an effective treatment nor a vaccine has been developed against CCHFV, surveillance programs are essential to limit and control the spread of the virus. In this study, a multiplex assay detecting antibodies against different CCHFV antigens in a single sample and independent of the ruminant species has been developed. This assay could be very useful in surveillance studies, to control the spread of CCHFV and prevent future outbreaks, and to better understand the immune response induced by CCHFV.
Disciplines :
Microbiology
Author, co-author :
Hoste, Alexis ; Université de Liège - ULiège > Département GxABT > Microbial technologies ; Eurofins-Inmunología y Genética Aplicada S.A. (Eurofins-INGENASA S.A.), Madrid, Spain ; School of Molecular and Cellular Biology, University of Leeds, Leeds, United Kingdom
Djadjovski, Igor; Ss. Cyril and Methodius University in Skopje, Faculty of Veterinary Medicine, Skopje, North Macedonia
Jiménez-Clavero, Miguel Ángel; Centro de Investigación en Sanidad Animal (CISA), Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA-CSIC), Valdeolmos, Spain ; CIBER of Epidemiology and Public Health (CIBERESP), Madrid, Spain
Multiplex Assay for Simultaneous Detection of Antibodies against Crimean-Congo Hemorrhagic Fever Virus Nucleocapsid Protein and Glycoproteins in Ruminants.
H2020 - 721367 - HONOURs - Host switching pathogens, infectious outbreaks and zoonosis; a Marie Sklodowska-Curie Training Network
Funders :
EU - European Union
Funding text :
We thank Isabel García and Mercedes Montón for technical assistance. We thank the EU Marie Skłodowska-Curie Actions (MSCA) Innovative Training Network (ITN): H2020- MSCA-ITN-2016, under grant no. 721367 (to Alexis C. R. Hoste) and the MediLabSecure Project, supported by the European Commission (DEVCO: IFS/2018/402-247) (to Igor Djadjovski and Miguel Ángel Jiménez-Clavero).
Lasecka L, Baron MD. 2014. The molecular biology of nairoviruses, an emerging group of tick-borne arboviruses. Arch Virol 159:1249–1265. https://doi.org/10.1007/s00705-013-1940-z.
Bente DA, Forrester NL, Watts DM, McAuley AJ, Whitehouse CA, Bray M. 2013. Crimean-Congo hemorrhagic fever: History, epidemiology, pathogenesis, clinical syndrome and genetic diversity. Antiviral Res 100:159–189. https://doi.org/10.1016/j.antiviral.2013.07.006.
Negredo A, Habela MÁ, de Arellano ER, Diez F, Lasala F, López P, Sarriá A, Labiod N, Calero-Bernal R, Arenas M, Tenorio A, Estrada-Peña A, Sánchez-Seco MP. 2019. Survey of Crimean-Congo hemorrhagic fever enzootic focus, Spain, 2011–2015. Emerg Infect Dis 25:1177–1184. https://doi.org/10.3201/eid2506.180877.
Maltezou HC, Papa A. 2010. Crimean-Congo hemorrhagic fever: Risk for emergence of new endemic foci in Europe? Travel Med Infect Dis 8:139–143. https://doi.org/10.1016/j.tmaid.2010.04.008.
Naderi HR, Sarvghad MR, Bojdy A, Hadizadeh MR, Sadeghi R, Sheybani F. 2011. Nosocomial outbreak of Crimean-Congo haemorrhagic fever. Epidemiol Infect 139:862–866. https://doi.org/10.1017/S0950268810002001.
International Organisation for Animal Health (OIE). 2018. Crimean-Congo hemorrhagic fever pp 399–406. In Terrestrial Manual, 8th ed. International Organisation for Animal Health (OIE), Paris, France.
Papa A, Mirazimi A, Köksal I, Estrada-Pena A, Feldmann H. 2015. Recent advances in research on Crimean-Congo hemorrhagic fever. J Clin Virol 64:137–143. https://doi.org/10.1016/j.jcv.2014.08.029.
Oestereich L, Rieger T, Neumann M, Bernreuther C, Lehmann M, Krasemann S, Wurr S, Emmerich P, de Lamballerie X, Ölschläger S, Günther S. 2014. Evaluation of antiviral efficacy of ribavirin, Arbidol, and T-705 (Favipiravir) in a mouse model for Crimean-Congo hemorrhagic fever. PLoS Negl Trop Dis 8: e2804. https://doi.org/10.1371/journal.pntd.0002804.
Hewson R, Chamberlain J, Mioulet V, Lloyd G, Jamil B, Hasan R, Gmyl A, Gmyl L, Smirnova SE, Lukashev A, Karganova G, Clegg C. 2004. Crimean-Congo haemorrhagic fever virus: sequence analysis of the small RNA segments from a collection of viruses world wide. Virus Res 102:185–189. https://doi.org/10.1016/j.virusres.2003.12.035.
Barnwal B, Karlberg H, Mirazimi A, Tan YJ. 2016. The non-structural protein of Crimean-Congo hemorrhagic fever virus disrupts the mitochondrial membrane potential and induces apoptosis. J Biol Chem 291:582–592. https://doi.org/10.1074/jbc.M115.667436.
Elliott RM. 1990. Molecular biology of the Bunyaviridae. J Gen Virol 71: 501–522. https://doi.org/10.1099/0022-1317-71-3-501.
Sas MA, Comtet L, Donnet F, Mertens M, Vatansever Z, Tordo N, Pourquier P, Groschup MH. 2018. A novel double-antigen sandwich ELISA for the species-independent detection of Crimean-Congo hemorrhagic fever virus-specific antibodies. Antiviral Res 151:24–26. https://doi.org/10.1016/j.antiviral.2018.01.006.
Shrivastava N, Shrivastava A, Ninawe SM, Sharma S, Kumar JS, Alam SI, Kanani A, Sharma SK, Dash PK. 2019. Development of multispecies recombinant nucleoprotein-based indirect ELISA for high-throughput screening of Crimean-Congo hemorrhagic fever virus-specific antibodies. Front Microbiol 10:1822. https://doi.org/10.3389/fmicb.2019.01822.
Gülce-Iz S, Elaldı N, Can H, S ahar EA, Karakavuk M, Gül A, Kumoğlu GÖ, Dös kaya AD, Gürüz AY, Özdarendeli A, Felgner PL, Davies H, Dös kaya M. 2021. Development of a novel recombinant ELISA for the detection of Crimean-Congo hemorrhagic fever virus IgG antibodies. Sci Rep 11:5936. https://doi.org/10.1038/s41598-021-85323-1.
Zivcec M, Scholte FEM, Spiropoulou CF, Spengler JR, Bergeron É. 2016. Molecular insights into Crimean-Congo hemorrhagic fever virus. Viruses 8:106. https://doi.org/10.3390/v8040106.
Sanchez AJ, Vincent MJ, Nichol ST. 2002. Characterization of the glycoproteins of Crimean-Congo hemorrhagic fever virus. J Virol 76:7263–7275. https://doi.org/10.1128/jvi.76.14.7263-7275.2002.
Sanchez AJ, Vincent MJ, Erickson BR, Nichol ST. 2006. Crimean-Congo hemorrhagic fever virus glycoprotein precursor is cleaved by furin-like and SKI-1 proteases to generate a novel 38-kilodalton glycoprotein. J Virol 80:514–525. https://doi.org/10.1128/JVI.80.1.514-525.2006.
Bertolotti-Ciarlet A, Smith J, Strecker K, Paragas J, Altamura LA, McFalls JM, Frias-Stäheli N, García-Sastre A, Schmaljohn CS, Doms RW. 2005. Cellular localization and antigenic characterization of Crimean-Congo hemorrhagic fever virus glycoproteins. J Virol 79:6152–6161. https://doi.org/10.1128/JVI.79.10.6152-6161.2005.
Bergeron É, Zivcec M, Chakrabarti AK, Nichol ST, Albariño CG, Spiropoulou CF. 2015. Recovery of recombinant Crimean Congo hemorrhagic fever virus reveals a function for non-structural glycoproteins cleavage by furin. PLoS Pathog 11:e1004879. https://doi.org/10.1371/journal.ppat.1004879.
Golden JW, Shoemaker CJ, Lindquist ME, Zeng X, Daye SP, Williams JA, Liu J, Coffin KM, Olschner S, Flusin O, Altamura LA, Kuehl KA, Fitzpatrick CJ, Schmaljohn CS, Garrison AR. 2019. GP38-targeting monoclonal antibodies protect adult mice against lethal Crimean-Congo hemorrhagic fever virus infection. Sci Adv 5:eaaw9535. https://doi.org/10.1126/sciadv.aaw9535.
Mishra A, Moyer C, Abelson D, Deer D, El Omari K, Duman R, Lobel L, Lutwama J, Dye J, Wagner A, Chandran K, Cross R, Geisbert T, Zeitlin L, Bornholdt Z, McLellan J. 2020. Structure and characterization of Crimean-Congo hemorrhagic fever virus GP38. J Virol 94:e02005-19. https://doi.org/10.1128/JVI.02005-19.
WHO. 2018. Roadmap for research and product development against Crimean-Congo haemorrhagic fever (CCHF). https://www.who.int/publications/m/item/crimean-congo-haemorrhagic-fever-(cchf)-research-and-development(r-d)-roadmap.
Estrada-Peña A, Ruiz-Fons F, Acevedo P, Gortazar C, de la Fuente J. 2013. Factors driving the circulation and possible expansion of Crimean-Congo haemorrhagic fever virus in the western Palearctic. J Appl Microbiol 114: 278–286. https://doi.org/10.1111/jam.12039.
Medlock JM, Leach SA. 2015. Effect of climate change on vector-borne disease risk in the UK. Lancet Infect Dis 15:721–730. https://doi.org/10.1016/S1473-3099(15)70091-5.
Walker PJ, Widen SG, Wood TG, Guzman H, Tesh RB, Vasilakis N. 2016. A global genomic characterization of nairoviruses identifies nine discrete genogroups with distinctive structural characteristics and host-vector associations. Am J Trop Med Hyg 94:1107–1122. https://doi.org/10.4269/ajtmh.15-0917.
van der Wal FJ, Achterberg RP, de Boer SM, Boshra H, Brun A, Maassen CBM, Kortekaas J. 2012. Bead-based suspension array for simultaneous detection of antibodies against the Rift Valley fever virus nucleocapsid and Gn glycoprotein. J Virol Methods 183:99–105. https://doi.org/10.1016/j.jviromet.2012.03.008.
Carter SD, Barr JN, Edwards TA. 2012. Expression, purification and crystallization of the Crimean-Congo haemorrhagic fever virus nucleocapsid protein. Acta Crystallogr Sect F Struct Biol Cryst Commun 68:569–573. https://doi.org/10.1107/S1744309112009736.
Rahpeyma M, Samarbaf-Zadeh A, Makvandi M, Ghadiri AA, Dowall SD, Fotouhi F. 2017. Expression and characterization of codon-optimized Crimean-Congo hemorrhagic fever virus Gn glycoprotein in insect cells. Arch Virol 162:1951–1962. https://doi.org/10.1007/s00705-017-3315-3.
Emmerich P, Mika A, von Possel R, Rackow A, Liu Y, Schmitz H, Günther S, Sherifi K, Halili B, Jakupi X, Berisha L, Ahmeti S, Deschermeier C. 2018. Sensitive and specific detection of Crimean-Congo hemorrhagic fever virus (CCHFV): specific IgM and IgG antibodies in human sera using recombinant CCHFV nucleoprotein as antigen in m-capture and IgG immune complex (IC) ELISA tests. PLoS Negl Trop Dis 12:e0006366. https://doi.org/10.1371/journal.pntd.0006366.
Surtees R, Stern D, Ahrens K, Kromarek N, Lander A, Kreher P, Weiss S, Hewson R, Punch EK, Barr JN, Witkowski PT, Couacy-Hymann E, Marzi A, Dorner BG, Kurth A. 2020. Development of a multiplex microsphere immunoassay for the detection of antibodies against highly pathogenic viruses in human and animal serum samples. PLoS Negl Trop Dis 14:e0008699. https://doi.org/10.1371/journal.pntd.0008699.
Hoste ACR, Ruiz T, Fernández-Pacheco P, Jiménez-Clavero MÁ, Djadjovski I, Moreno S, Brun A, Edwards TA, Barr JN, Rueda P, Sastre P. 2020. Development of a multiplex assay for antibody detection in serum against pathogens affecting ruminants. Transbound Emerg Dis 68:1229–1239. https://doi.org/10.1111/tbed.13776.
Hurtado A, Rueda P, Nowicky J, Sarraseca J, Casal JI. 1996. Identification of domains in canine parvovirus VP2 essential for the assembly of virus-like particles. J Virol 70:5422–5429. https://doi.org/10.1128/JVI.70.8.5422-5429.1996.
Angeloni S, Cordes R, Dunbar S, Garcia C, Gibson G, Martin C, Stone V. 2016. xMAP Cookbook, 3rd ed. Luminex Corporation, Austin, Texas, USA.
Hermanson GT. 2013. Zero-length crosslinkers, p 259–273. In Bioconjugate techniques, 3rd ed. Academic Press, Cambridge, Massachusetts, USA.
