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Journal of Medical Microbiology logo Journal of Medical Microbiology

Volume 73, Issue 6

Detection of avian reovirus (ARV) by ELISA based on recombinant σB, σC and σNS full-length proteins and protein fragments No Access

Abstract

Avian reovirus (ARV) is associated with arthritis/tenosynovitis and malabsorption syndrome in chickens. The σC and σB proteins, both exposed to the virus capsid, are highly immunogenic and could form the basis for diagnostic devices designed to assess the immunological status of the flock.

Commercial ARV ELISAs cannot distinguish between vaccinated and infected animals and might not detect circulating ARV strains.

We aimed to develop a customized test to detect the circulating field ARV strains as well as distinguish between vaccinated and unvaccinated animals.

We developed ELISA assays based on recombinant (r) σB, σC and the nonstructural protein σNS and tested them using antisera of vaccinated and unvaccinated chickens as well as negative controls. Fragments of σB and σC proteins were also used to study regions that could be further exploited in diagnostic tests.

Vaccinated and unvaccinated birds were positive by commercial ELISA, with no difference in optical density values. In contrast, samples of unvaccinated animals showed lower absorbance in the rσB and rσC ELISA tests and higher absorbance in the rσNS ELISA test than the vaccinated animals. Negative control samples were negative in all tests. Fragmentation of σB and σC proteins showed that some regions can differentiate between vaccinated and unvaccinated animals. For example, σB amino acids 128–179 (σB-F4) and σC amino acids 121–165 (σC-F4) exhibited 85 and 95% positivity among samples of vaccinated animals but only 5% and zero positivity among samples of unvaccinated animals, respectively.

These data suggest that unvaccinated birds might have been exposed to field strains of ARV. The reduction in absorbance in the recombinant tests possibly reflects an increased specificity of our test since unvaccinated samples showed less cross-reactivity with the vaccine proteins immobilized on ELISAs. The discrepant results obtained with the protein fragment tests between vaccinated and unvaccinated animals are discussed in light of the diversity between ARV strains.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): antigenic regions , avian reovirus , diagnosis and ELISA
Funding
This study was supported by the:
  • Conselho Nacional de Desenvolvimento Científico e Tecnológico
    • Principle Award Recipient: TatianaReichert Assunção de Matos
  • Instituto de Biologia Molecular do Paraná
    • Principle Award Recipient: NotApplicable
  • Instituto Carlos Chagas
    • Principle Award Recipient: NotApplicable
© 2024 The Authors
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2024-06-27
2025-04-26
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References

  1. Van Der Heide L. Viral arthritis/tenosynovitis: a review. Avian Pathol 1977; 6:271–284 [View Article] [PubMed]
     [Google Scholar]
  2. Jones RC. Avian reovirus infections. Rev Sci Tech Oie 2000; 19:614–625 [View Article]
     [Google Scholar]
  3. Menendez NA, Calnek BW, Cowen BS. Experimental egg-transmission of avian reovirus. Avian Dis 1975; 19:104–111 [PubMed]
     [Google Scholar]
  4. van der Heide L, Kalbac M. Infectious tenosynovitis (viral arthritis): characterization of a Connecticut viral isolant as a reovirus and evidence of viral egg transmission by reovirus-infected broiler breeders. Avian Dis 1975; 19:683–688 [PubMed]
     [Google Scholar]
  5. al Afaleq AI, Jones RC. Localisation of avian reovirus in the hock joints of chicks after entry through broken skin. Res Vet Sci 1990; 48:381–382 [PubMed]
     [Google Scholar]
  6. Spandidos DA, Graham AF. Physical and chemical characterization of an avian reovirus. J Virol 1976; 19:968–976 [View Article] [PubMed]
     [Google Scholar]
  7. Zhang X, Tang J, Walker SB, O’Hara D, Nibert ML et al. Structure of avian orthoreovirus virion by electron cryomicroscopy and image reconstruction. Virology 2005; 343:25–35 [View Article]
     [Google Scholar]
  8. Schnitzer TJ, Ramos T, Gouvea V. Avian reovirus polypeptides: analysis of intracellular virus-specified products, virions, top component, and cores. J Virol 1982; 43:1006–1014 [View Article] [PubMed]
     [Google Scholar]
  9. Ni Y, Ramig RF, Kemp MC. Identification of proteins encoded by avian reoviruses and evidence for post-translational modification. Virology 1993; 193:466–469 [View Article] [PubMed]
     [Google Scholar]
  10. Wickramasinghe R, Meanger J, Enriquez CE, Wilcox GE. Avian reovirus proteins associated with neutralization of virus infectivity. Virology 1993; 194:688–696 [View Article] [PubMed]
     [Google Scholar]
  11. Varela R, Benavente J. Protein coding assignment of avian reovirus strain S1133. J Virol 1994; 68:6775–6777 [View Article] [PubMed]
     [Google Scholar]
  12. Shmulevitz M, Duncan R. A new class of fusion-associated small transmembrane (FAST) proteins encoded by the non-enveloped fusogenic reoviruses. EMBO J 2000; 19:902–912 [View Article] [PubMed]
     [Google Scholar]
  13. Bodelón G, Labrada L, Martínez-Costas J, Benavente J. The avian reovirus genome segment S1 is a functionally tricistronic gene that expresses one structural and two nonstructural proteins in infected cells. Virology 2001; 290:181–191 [View Article] [PubMed]
     [Google Scholar]
  14. Benavente J, Martínez-Costas J. Avian reovirus: structure and biology. Virus Res 2007; 123:105–119 [View Article] [PubMed]
     [Google Scholar]
  15. Kant A, Balk F, Born L, van Roozelaar D, Heijmans J et al. Classification of Dutch and German avian reoviruses by sequencing the sigma C protein. Vet Res 2003; 34:203–212 [View Article] [PubMed]
     [Google Scholar]
  16. Liu HJ, Lee LH, Hsu HW, Kuo LC, Liao MH. Molecular evolution of avian reovirus: evidence for genetic diversity and reassortment of the S-class genome segments and multiple cocirculating lineages. Virology 2003; 314:336–349 [View Article] [PubMed]
     [Google Scholar]
  17. Liu HJ, Lee LH, Shih WL, Li YJ, Su HY. Rapid characterization of avian reoviruses using phylogenetic analysis, reverse transcription-polymerase chain reaction and restriction enzyme fragment length polymorphism. Avian Pathol 2004; 33:171–180 [View Article] [PubMed]
     [Google Scholar]
  18. Hsu HW, Su HY, Huang PH, Lee BLH, Liu HJ. Sequence and phylogenetic analysis of P10- and P17-encoding genes of avian reovirus. Avian Dis 2005; 49:36–42 [View Article] [PubMed]
     [Google Scholar]
  19. Shen PC, Chiou YF, Liu HJ, Song CH, Su YP et al. Genetic variation of the lambdaA and lambdaC protein encoding genes of avian reoviruses. Res Vet Sci 2007; 83:394–402 [View Article] [PubMed]
     [Google Scholar]
  20. Goldenberg D, Pasmanik-Chor M, Pirak M, Kass N, Lublin A et al. Genetic and antigenic characterization of sigma C protein from avian reovirus. Avian Pathol 2010; 39:189–199 [View Article] [PubMed]
     [Google Scholar]
  21. Teng L, Xie Z, Xie L, Liu J, Pang Y et al. Sequencing and phylogenetic analysis of an avian reovirus genome. Virus Genes 2014; 48:381–386 [View Article] [PubMed]
     [Google Scholar]
  22. Lu H, Tang Y, Dunn PA, Wallner-Pendleton EA, Lin L et al. Isolation and molecular characterization of newly emerging avian reovirus variants and novel strains in Pennsylvania, USA, 2011–2014. Sci Rep 2011; 5: [View Article] [PubMed]
     [Google Scholar]
  23. De la Torre D, Astolfi-Ferreira CS, Chacón RD, Puga B, Piantino Ferreira AJ. Emerging new avian reovirus variants from cases of enteric disorders and arthritis/tenosynovitis in Brazilian poultry flocks. Br Poult Sci 2021; 62:361–372 [View Article] [PubMed]
     [Google Scholar]
  24. Jones RC, Georgiou K. Reovirus-induced tenosynovitis in chickens the influence of age at infection. Avian Pathol 1984; 13:441–457 [View Article] [PubMed]
     [Google Scholar]
  25. van der Heide L. The history of avian reovirus. Avian Dis 2000; 44:638–641 [PubMed]
     [Google Scholar]
  26. Vasserman Y, Eliahoo D, Hemsani E, Kass N, Ayali G et al. The influence of reovirus sigma C protein diversity on vaccination efficiency. Avian Dis 2004; 48:271–278 [View Article] [PubMed]
     [Google Scholar]
  27. Goldenberg D, Lublin A, Rosenbluth E, Heller ED, Pitcovski J. Differentiating infected from vaccinated animals, and among virulent prototypes of reovirus. J Virol Methods 2011; 177:80–86 [View Article] [PubMed]
     [Google Scholar]
  28. Lublin A, Goldenberg D, Rosenbluth E, Heller ED, Pitcovski J. Wide-range protection against avian reovirus conferred by vaccination with representatives of four defined genotypes. Vaccine 2011; 29:8683–8688 [View Article] [PubMed]
     [Google Scholar]
  29. Davis JF, Kulkarni A, Fletcher O. Reovirus infections in young broiler chickens. Avian Dis 2013; 57:321–325 [View Article] [PubMed]
     [Google Scholar]
  30. Troxler S, Rigomier P, Bilic I, Liebhart D, Prokofieva I et al. Identification of a new reovirus causing substantial losses in broiler production in France, despite routine vaccination of breeders. Vet Rec 2013; 172:556 [View Article] [PubMed]
     [Google Scholar]
  31. Souza SO, De Carli S, Lunge VR, Ikuta N, Canal CW et al. Pathological and molecular findings of avian reoviruses from clinical cases of tenosynovitis in poultry flocks from Brazil. Poult Sci 2018; 97:3550–3555 [View Article] [PubMed]
     [Google Scholar]
  32. De Carli S, Wolf JM, Gräf T, Lehmann FKM, Fonseca ASK et al. Genotypic characterization and molecular evolution of avian reovirus in poultry flocks from Brazil. Avian Pathol 2020; 49:611–620 [View Article] [PubMed]
     [Google Scholar]
  33. Shapouri MRS, Frenette D, Larochelle R, Arella M, Silim A. Characterization of monoclonal antibodies against avian reovirus strain S1133. Avian Pathol 1996; 25:57–67 [View Article] [PubMed]
     [Google Scholar]
  34. Lin YH, Lee LH, Shih WL, Hu YC, Liu HJ. Baculovirus surface display of sigmaC and sigmaB proteins of avian reovirus and immunogenicity of the displayed proteins in a mouse model. Vaccine 2008; 26:6361–6367 [View Article] [PubMed]
     [Google Scholar]
  35. Grande A, Rodriguez E, Costas C, Everitt E, Benavente J. Oligomerization and cell-binding properties of the avian reovirus cell-attachment protein sigmaC. Virology 2000; 274:367–377 [View Article] [PubMed]
     [Google Scholar]
  36. Guardado Calvo P, Fox GC, Hermo Parrado XL, Llamas-Saiz AL, Costas C et al. Structure of the carboxy-terminal receptor-binding domain of avian reovirus fibre sigmaC. J Mol Biol 2005; 354:137–149 [View Article] [PubMed]
     [Google Scholar]
  37. Shapouri MRS, Arella M, Silim A. Evidence for the multimeric nature and cell binding ability of avian reovirus3 protein. J Gen Virol 1996; 77:1203–1210 [View Article]
     [Google Scholar]
  38. Martínez-Costas J, Grande A, Varela R, García-Martínez C, Benavente J. Protein architecture of avian reovirus S1133 and identification of the cell attachment protein. J Virol 1997; 71:59–64 [View Article]
     [Google Scholar]
  39. Guardado-Calvo P, Fox GC, Llamas-Saiz AL, van Raaij MJ. Crystallographic structure of the alpha-helical triple coiled-coil domain of avian reovirus S1133 fibre. J Gen Virol 2009; 90:672–677 [View Article] [PubMed]
     [Google Scholar]
  40. Yin HS, Lee LH. Identification and characterization of RNA-binding activities of avian reovirus non-structural protein sigmaNS. J Gen Virol 1998; 79 (Pt 6):1411–1413 [View Article] [PubMed]
     [Google Scholar]
  41. Borodavka A, Ault J, Stockley PG, Tuma R. Evidence that avian reovirus σNS is an RNA chaperone: implications for genome segment assortment. Nucleic Acids Res 2015; 43:7044–7057 [View Article] [PubMed]
     [Google Scholar]
  42. Xie Z, Qin C, Xie L, Liu J, Pang Y et al. Recombinant protein-based ELISA for detection and differentiation of antibodies against avian reovirus in vaccinated and non-vaccinated chickens. J Virol Methods 2010; 165:108–111 [View Article] [PubMed]
     [Google Scholar]
  43. Shien JH, Yin HS, Lee LH. An enzyme-linked immunosorbent assay for the detection of antibody to avian reovirus by using protein sigma B as the coating antigen. Res Vet Sci 2000; 69:107–112 [View Article] [PubMed]
     [Google Scholar]
  44. Liu HJ, Kuo LC, Hu YC, Liao MH, Lien YY. Development of an ELISA for detection of antibodies to avian reovirus in chickens. J Virol Methods 2002; 102:129–138 [View Article] [PubMed]
     [Google Scholar]
  45. Yang Z-J, Wang C-Y, Lee L-H, Chuang K-P, Lien Y-Y et al. Development of ELISA kits for antibodies against avian reovirus using the sigmaC and sigmaB proteins expressed in the methyltropic yeast Pichia pastoris. J Virol Methods 2010; 163:169–174 [View Article] [PubMed]
     [Google Scholar]
  46. Hou HS, Su YP, Shieh HK, Lee LH. Monoclonal antibodies against different epitopes of nonstructural protein sigmaNS of avian reovirus S1133. Virology 2001; 282:168–175 [View Article] [PubMed]
     [Google Scholar]
  47. Hsu CJ, Wang CY, Lee LH, Shih WL, Chang CI et al. Development and characterization of monoclonal antibodies against avian reovirus sigma C protein and their application in detection of avian reovirus isolates. Avian Pathol 2006; 35:320–326 [View Article] [PubMed]
     [Google Scholar]
  48. Yin C, Qin L, Sun M, Gao Y, Qi X et al. Identification of a linear B-Cell epitope on avian reovirus protein sigmaC. Virus Res 2013; 178:530–534 [View Article] [PubMed]
     [Google Scholar]
  49. Yin C, Qin L, Sun M, Gao Y, Qi X et al. Antigenic analysis of monoclonal antibodies against different epitopes of σB protein of avian reovirus. PLoS One 2013; 8:e81533 [View Article] [PubMed]
     [Google Scholar]
  50. Huang PH, Li YJ, Su YP, Lee LH, Liu HJ. Epitope mapping and functional analysis of sigma A and sigma NS proteins of avian reovirus. Virology 2005; 332:584–595 [View Article] [PubMed]
     [Google Scholar]
  51. Kyte J, Doolittle RF. A simple method for displaying the hydropathic character of a protein. J Mol Biol 1982; 157:105–132 [View Article] [PubMed]
     [Google Scholar]
  52. Emini EA, Hughes JV, Perlow DS, Boger J. Induction of hepatitis A virus-neutralizing antibody by a virus-specific synthetic peptide. J Virol 1985; 55:836–839 [View Article] [PubMed]
     [Google Scholar]
  53. Jameson BA, Wolf H. The antigenic index: a novel algorithm for predicting antigenic determinants. Bioinformatics 1988; 4:181–186 [View Article]
     [Google Scholar]
  54. Aoki T, Kaneta M, Onagi H, Takahashi Y, Koch KS et al. A simple and rapid immunoassay system using green fluorescent protein tag. J Immunoassay 1997; 18:321–333 [View Article] [PubMed]
     [Google Scholar]
  55. Clark FD, Ni Y, Collisson EW, Kemp MC. Characterization of avian reovirus strain-specific polymorphisms. Avian Dis 1990; 34:304–314 [PubMed]
     [Google Scholar]
  56. Gallardo RA. Molecular characterization of variant avian reoviruses and their relationship with antigenicity and pathogenicity. Avian Dis 2022; 66:443–446 [View Article] [PubMed]
     [Google Scholar]
  57. Nour I, Alvarez-Narvaez S, Harrell TL, Conrad SJ, Mohanty SK. Whole genomic constellation of avian reovirus strains isolated from broilers with arthritis in North Carolina, USA. Viruses 2023; 15:2191 [View Article]
     [Google Scholar]
  58. Goldenberg D, Lublin A, Rosenbluth E, Heller ED, Pitcovski J. Optimized polypeptide for a subunit vaccine against avian reovirus. Vaccine 2016; 34:3178–3183 [View Article] [PubMed]
     [Google Scholar]
  59. Sun Z, Zhu Y, Wang Q, Ye L, Dai Y et al. An immunogen containing four tandem 10E8 epitope repeats with exposed key residues induces antibodies that neutralize HIV-1 and activates an ADCC reporter gene. Emerg Microbes Infect 2016; 5:e65 [View Article] [PubMed]
     [Google Scholar]
  60. Qin Y, Tu K, Teng Q, Feng D, Zhao Y et al. Identification of novel T-cell epitopes on infectious bronchitis virus N protein and development of a multi-epitope vaccine. J Virol 2021; 95:e0066721 [View Article]
     [Google Scholar]
  61. Dawe WH, Kapczynski DR, Linnemann EG, Gauthiersloan VR, Sellers HS. Analysis of the immune response and identification of antibody epitopes against the sigma C protein of avian orthoreovirus following immunization with live or inactivated vaccines. Avian Dis 2022; 66:465–478 [View Article] [PubMed]
     [Google Scholar]
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Detection of avian reovirus (ARV) by ELISA based on recombinant σB, σC and σNS full-length proteins and protein fragments
J. Med. Microbiol. 73, 001836 (2024); https://doi.org/10.1099/jmm.0.001836
/content/journal/jmm/10.1099/jmm.0.001836
/content/journal/jmm/10.1099/jmm.0.001836
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