Abstract
The Bluetongue virus (BTV) VP7 protein represents an important group-specific antigen that can serve as a basis for diagnostic tests. Here, we report the generation of a novel BTV group-specific monoclonal antibody (Mab; herein named 4H7) that recognizes a conformational epitope in the VP7 protein. We used a phage-displayed peptide screen and site-directed mutagenesis to define the VP7 amino acid residues that most strongly contribute to the conformational epitope recognized by Mab 4H7. Amino acid residues at positions 175, 185, 186, and 278 of the BTV VP7 protein strongly contributed to Mab 4H7 binding. These key amino acid residues are conserved among all BTV serotypes, whereas related Orbiviruses possess at least one amino acid substitution at these positions. We developed a competitive enzyme-linked immunosorbent assay (c-ELISA) using Mab 4H7 and recombinant BTV VP7 protein to detect serum antibodies against this BTV group-specific VP7 epitope. The c-ELISA was used to screen 833 clinical samples collected from animals in three provinces of China. BTV seroprevalence in the three provinces ranged from 25.42 to 47.45 %. This work provides the foundation for a new diagnostic c-ELISA that can be further applied to BTV surveillance activities and informs our understanding of the structure of the BTV VP7 protein.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Afshar A, Thomas FC, Wright PF, Shapiro JL, Anderson J (1989) Comparison of competitive ELISA, indirect ELISA and standard AGID tests for detecting blue-tongue virus antibodies in cattle and sheep. Vet Rec 124(6):136–141
Brocks B, Garin-Chesa P, Behrle E, Park JE, Retting WJ, Pfizenmaier K, Moosmayer D (2001) Species-crossreactive scFv against the tumor stroma marker “fibroblast activation protein” selected by phage display from an immunized FAP-/- knock-out mouse. Mol Med 7(7):461–469
du Plessis DH, Wang LF, Jordaan FA, Eaton BT (1994) Fine mapping of a continuous epitope on VP7 of bluetongue virus using overlapping synthetic peptides and a random epitope library. Virology 198(1):346–349
Elbers AR, Backx A, Mintiens K, Gerbier G, Staubach C, Hendrickx G, van der Spek A (2008) Field observation during the Bluetongue serotype 8 epidemic in 2006. II. Morbidity and mortality rate, case fatality and clinical recovery in sheep and cattle in the Netherlands. Prev Vet Med 87(1–2):31–40. doi:10.1016/j
GENG H-w, QIN Y-l, LI J-p, Yang T, Sun E-c, Liu N-h, Bai A-b, Xu Q-y, Wang L-f, Zhao J, Qin S-m, Lin J, Guo Y-f, Wu D-l (2012) Development of monoclone antibody competitive ELISA for detection of bluetongue virus antibodies. Chin J Prev Vet Med 34(5):388–392
Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256(5517):495–497
Li JK, Yang YY (1990) Mapping of two immunodominant antigenic epitopes conserved among the major inner capsid protein, VP7 of five bluetongue viruses. Virology 178(2):552–559
Martyn JC, Gould AR, Eaton BT (1991) High level expression of the major core protein VP7 and the non-structural protein NS3 of bluetongue virus in yeast: use of expressed VP7 as a diagnostic, group-reactive antigen in a blocking ELISA. Virus Res 18(2-3):165–178. doi:10.1128/jcm.42.2.518-523.2004
Mecham JO, Wilson WC (2004) Antigen capture competitive enzyme-linked immunosorbent assay using baculovirus-expressed antigens for diagnosis of bluetongue virus and epizootic hemorrhagic disease virus. J Clin Microbiol 42(2):518–523. doi:10.1128/jcm.42.2.518-523.2004
Mertens PP, Maan NS, Prasad G, Samuel AR, Shaw AE, Potgieter AC, Anthony SJ, Maan S (2007) Design of primers and use of RT-PCR assays for typing European bluetongue virus isolates: differentiation of field and vaccine strains. J Gen Virol 88(Pt10):2811–2823. doi:10.1099/vir.0.83023-0
Maan S, Maan NS, Nomikou K, Veronesi E, Bachanek-Bankowska K, Belaganahalli MN, Attoui H, Mertens PP (2011) Complete genome characterisation of a novel 26th bluetongue virus serotype from Kuwait. PLoS One 6(10):e26147. doi:10.1371/journal.pone.0026147
Reddington JJ, Reddington GM, MacLachlan NJ (1991) A competitive ELISA for detection of antibodies to the group antigen of bluetongue virus. J Vet Diagn Investig 3(2):144–147
Ratinier M, Caporale M, Golder M, Franzoni G, Allan K, Nunes SF, Armezzani A, Bayoumy A, Rixon F, Shaw A, Palmarini M (2011) Identification and characterization of a novel non-structural protein of bluetongue virus. PLoS Pathog 7(12):e1002477. doi:10.1371/journal.ppat.1002477
Sun EC, Zhao J, Liu NH, Yang T, Ma JN, Geng HW, Wang LF, Qin YL, Bu ZG, Yang YH, Lunt RA, Wang LF, Wu DL (2012) Comprehensive mapping of West Nile virus (WNV)- and Japanese encephalitis virus serocomplex-specific linear B-cell epitopes from WNV non-structural protein 1. J Gen Virol 93(Pt):50–60. doi:10.1099/vir.0.034900-0
Vögtlin A, Hofmann MA, Nenniger C, Renzullo S, Steinrigl A, Loitsch A, Schwermer H, Kaufmann C, Thür B (2013) Long-term infection of goats with bluetongue virus serotype 25. Vet Microbiol 166(1–2):165–173. doi:10.1016/j.vetmic.2013.06.001
Wang LF, Scanlon DB, Kattenbelt JA, Mecham JO, Eaton BT (1994) Fine mapping of a surface-accessible, immunodominant site on the bluetongue virus major core protein VP7. Virology 204(2):811–814
Wang LF, Hyatt AD, Whiteley PL, Andrew M, Li JK, Eaton BT (1996) Topography and immunogenicity of bluetongue virus VP7 epitopes. Arch Virol 141(1):111–123
Wang WS, Sun EC, Liu NH, Yang T, Xu QY, Qin YL, Zhao J, Feng YF, Li JP, Wei P, Zhang CY, Wu DL (2012) Monoclonal antibodies against VP7 of bluetongue virus. Hybridomas (Larchmt) 31(6):469–472. doi:10.1089/hyb.2012.0046
Yang J, Hua Q, Chen H, Lv J, Chen B, Ruan Z (2010) A rapid assay for detecting antibody against Bluetongue virus with a latex agglutination test using recombinant VP7 antigen. J Vet Diagn Investig 22(2):242–244. doi:10.1177/104063871002200212
Zhang N, Li Z, Zhang F, Zhu J (2004) Studies on bluetongue disease in the People’s Republic of China. Vet Ital 40(3):51–56
Acknowledgments
This study was supported by the National Natural Science Foundation of China (grant no. 31302065), the China Postdoctoral Science Foundation (2013 M540172), Special Fund for Agroscientific Research in the Public Interest (201203056), and the State Key Laboratory of Veterinary Biotechnology (SKLVBP201313 and 201316). The authors thank Dr. Peter Wilker for editing the manuscript.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
ESM 1
(PDF 10425 kb)
Rights and permissions
About this article
Cite this article
Xu, Q.Y., Sun, E.C., Yang, T. et al. Isolation of a Bluetongue virus group-specific monoclonal antibody and application to a diagnostic competitive ELISA. Appl Microbiol Biotechnol 99, 729–739 (2015). https://doi.org/10.1007/s00253-014-5937-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00253-014-5937-7