1887

Abstract

Both porcine epidemic diarrhoea virus (PEDV) and porcine transmissible gastroenteritis virus (TGEV), which cause high mortality in piglets and produce similar clinical symptoms and histopathological morphology, belong to the genus Alphacoronavirus. Serological diagnosis plays an important role in distinguishing pathogen species. Together with the spike (S) protein, the nucleocapsid (N) protein is one of the immunodominant regions among coronaviruses. In this study, two-way antigenic cross-reactivity between the N proteins of PEDV and TGEV was observed by indirect immunofluorescence assay (IFA) and Western blot analysis. Furthermore, the PEDV N protein harbouring truncations of amino acids (aa) 1 to 170 or aa 125 to 301 was demonstrated to cross-react with the anti-TGEV N polyclonal antibody (PAb), whereas the truncation-expressing aa 302 to 401 resulted in a specific reaction with the anti-PEDV N PAb but not with the anti-TGEV N PAb. Mutants of the PEDV N protein were generated based on sequence alignment and structural analysis; we then confirmed that the N-terminal residues 58-RWRMRRGERIE-68 and 78-LGTGPHAD-85 contributed to the cross-reactivity. All the results provide vital clues for the development of precise diagnostic assays for porcine coronaviruses.

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2019-01-17
2019-09-18
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References

  1. Stevenson GW, Hoang H, Schwartz KJ, Burrough ER, Sun D et al. Emergence of Porcine epidemic diarrhea virus in the United States: clinical signs, lesions, and viral genomic sequences. J Vet Diagn Invest 2013;25:649–654 [CrossRef][PubMed]
    [Google Scholar]
  2. Cima G. Fighting a deadly pig disease. Industry, veterinarians trying to contain PED virus, new to the US. J Am Vet Med Assoc 2013;243:469–470[PubMed]
    [Google Scholar]
  3. Song D, Park B. Porcine epidemic diarrhoea virus: a comprehensive review of molecular epidemiology, diagnosis, and vaccines. Virus Genes 2012;44:167–175 [CrossRef][PubMed]
    [Google Scholar]
  4. Jung K, Saif LJ. Porcine epidemic diarrhea virus infection: Etiology, epidemiology, pathogenesis and immunoprophylaxis. Vet J 2015;204:134–143 [CrossRef][PubMed]
    [Google Scholar]
  5. Saif LJ. Coronavirus immunogens. Vet Microbiol 1993;37:285–297 [CrossRef][PubMed]
    [Google Scholar]
  6. Adams MJ, Carstens EB. Ratification vote on taxonomic proposals to the international committee on taxonomy of viruses (2012). Arch Virol 2012;157:1411–1422 [CrossRef][PubMed]
    [Google Scholar]
  7. Jung K, Wang Q, Scheuer KA, Lu Z, Zhang Y et al. Pathology of US porcine epidemic diarrhea virus strain PC21A in gnotobiotic pigs. Emerg Infect Dis 2014;20:668–671 [CrossRef][PubMed]
    [Google Scholar]
  8. Thomas JT, Chen Q, Gauger PC, Giménez-Lirola LG, Sinha A et al. Effect of porcine epidemic diarrhea virus infectious doses on infection outcomes in naïve conventional neonatal and weaned pigs. PLoS One 2015;10:e0139266 [CrossRef][PubMed]
    [Google Scholar]
  9. Saif LJ, Pensaert MB, Sestak K, Yeo SG, Jung K et al. Coronaviruses. In Zimmerman J, Karriker L, Ramirez A, Schwartz K, Stevenson G et al. (editors) Disease of Swine Ames, IA: Wiley-Blackwell; pp.501–524
    [Google Scholar]
  10. Kim SH, Kim IJ, Pyo HM, Tark DS, Song JY et al. Multiplex real-time RT-PCR for the simultaneous detection and quantification of transmissible gastroenteritis virus and porcine epidemic diarrhea virus. J Virol Methods 2007;146:172–177 [CrossRef][PubMed]
    [Google Scholar]
  11. Song DS, Kang BK, Oh JS, Ha GW, Yang JS et al. Multiplex reverse transcription-PCR for rapid differential detection of porcine epidemic diarrhea virus, transmissible gastroenteritis virus, and porcine group A rotavirus. J Vet Diagn Invest 2006;18:278–281 [CrossRef][PubMed]
    [Google Scholar]
  12. Okda F, Liu X, Singrey A, Clement T, Nelson J et al. Development of an indirect ELISA, blocking ELISA, fluorescent microsphere immunoassay and fluorescent focus neutralization assay for serologic evaluation of exposure to North American strains of porcine epidemic Diarrhea virus. BMC Vet Res 2015;11:180 [CrossRef][PubMed]
    [Google Scholar]
  13. Hou XL, Yu LY, Liu J. Development and evaluation of enzyme-linked immunosorbent assay based on recombinant nucleocapsid protein for detection of porcine epidemic diarrhea (PEDV) antibodies. Vet Microbiol 2007;123:86–92 [CrossRef][PubMed]
    [Google Scholar]
  14. Song Y, Singh P, Nelson E, Ramamoorthy S. A computationally designed serological assay for porcine epidemic diarrhea virus. J Clin Microbiol 2016;54:2039–2046 [CrossRef][PubMed]
    [Google Scholar]
  15. Sturman LS, Holmes KV. The molecular biology of coronaviruses. Adv Virus Res 1983;28:35–112[PubMed]
    [Google Scholar]
  16. Lin CM, Gao X, Oka T, Vlasova AN, Esseili MA et al. Antigenic relationships among porcine epidemic diarrhea virus and transmissible gastroenteritis virus strains. J Virol 2015;89:3332–3342 [CrossRef][PubMed]
    [Google Scholar]
  17. Gimenez-Lirola LG, Zhang J, Carrillo-Avila JA, Chen Q, Magtoto R et al. Reactivity of porcine epidemic Diarrhea virus structural proteins to antibodies against porcine enteric coronaviruses: diagnostic implications. J Clin Microbiol 2017;55:1426–1436 [CrossRef][PubMed]
    [Google Scholar]
  18. Wang K, Xie C, Zhang J, Zhang W, Yang D et al. The identification and characterization of two novel epitopes on the nucleocapsid protein of the porcine epidemic diarrhea virus. Sci Rep 2016;6:39010 [CrossRef][PubMed]
    [Google Scholar]
  19. Martín Alonso JM, Balbín M, Garwes DJ, Enjuanes L, Gascón S et al. Antigenic structure of transmissible gastroenteritis virus nucleoprotein. Virology 1992;188:168–174 [CrossRef][PubMed]
    [Google Scholar]
  20. Simkins RA, Saif LJ, Weilnau PA. Epitope mapping and the detection of transmissible gastroenteritis viral proteins in cell culture using biotinylated monoclonal antibodies in a fixed-cell ELISA. Arch Virol 1989;107:179–190 [CrossRef][PubMed]
    [Google Scholar]
  21. Ma Y, Zhang Y, Liang X, Oglesbee M, Krakowka S et al. Two-way antigenic cross-reactivity between porcine epidemic diarrhea virus and porcine deltacoronavirus. Vet Microbiol 2016;186:90–96 [CrossRef][PubMed]
    [Google Scholar]
  22. Horzinek MC, Lutz H, Pedersen NC. Antigenic relationships among homologous structural polypeptides of porcine, feline, and canine coronaviruses. Infect Immun 1982;37:1148–1155[PubMed]
    [Google Scholar]
  23. Vlasova AN, Zhang X, Hasoksuz M, Nagesha HS, Haynes LM et al. Two-way antigenic cross-reactivity between severe acute respiratory syndrome coronavirus (SARS-CoV) and group 1 animal CoVs is mediated through an antigenic site in the N-terminal region of the SARS-CoV nucleoprotein. J Virol 2007;81:13365–13377 [CrossRef][PubMed]
    [Google Scholar]
  24. Zhou YL, Ederveen J, Egberink H, Pensaert M, Horzinek MC. Porcine epidemic diarrhea virus (CV 777) and feline infectious peritonitis virus (FIPV) are antigenically related. Arch Virol 1988;102:63–71[PubMed]
    [Google Scholar]
  25. Have P, Moving V, Svansson V, Uttenthal A, Bloch B. Coronavirus infection in mink (Mustela vison). Serological evidence of infection with a coronavirus related to transmissible gastroenteritis virus and porcine epidemic diarrhea virus. Vet Microbiol 1992;31:1–10 [CrossRef][PubMed]
    [Google Scholar]
  26. He Y, Zhou Y, Wu H, Kou Z, Liu S et al. Mapping of antigenic sites on the nucleocapsid protein of the severe acute respiratory syndrome coronavirus. J Clin Microbiol 2004;42:5309–5314 [CrossRef][PubMed]
    [Google Scholar]
  27. Liang Y, Wan Y, Qiu LW, Zhou J, Ni B et al. Comprehensive antibody epitope mapping of the nucleocapsid protein of severe acute respiratory syndrome (SARS) coronavirus: insight into the humoral immunity of SARS. Clin Chem 2005;51:1382–1396 [CrossRef][PubMed]
    [Google Scholar]
  28. Huang Q, Yu L, Petros AM, Gunasekera A, Liu Z et al. Structure of the N-terminal RNA-binding domain of the SARS CoV nucleocapsid protein. Biochemistry 2004;43:6059–6063 [CrossRef][PubMed]
    [Google Scholar]
  29. Takeda M, Chang CK, Ikeya T, Güntert P, Chang YH et al. Solution structure of the c-terminal dimerization domain of SARS coronavirus nucleocapsid protein solved by the SAIL-NMR method. J Mol Biol 2008;380:608–622 [CrossRef][PubMed]
    [Google Scholar]
  30. Wang YS, Chang CK, Hou MH. Crystallographic analysis of the N-terminal domain of middle east respiratory syndrome coronavirus nucleocapsid protein. Acta Crystallogr F Struct Biol Commun 2015;71:977–980 [CrossRef][PubMed]
    [Google Scholar]
  31. Jayaram H, Fan H, Bowman BR, Ooi A, Jayaram J et al. X-ray structures of the N- and C-terminal domains of a coronavirus nucleocapsid protein: implications for nucleocapsid formation. J Virol 2006;80:6612–6620 [CrossRef][PubMed]
    [Google Scholar]
  32. Szelazek B, Kabala W, Kus K, Zdzalik M, Twarda-Clapa A et al. Structural characterization of human coronavirus NL63 N protein. J Virol 2017;91: [CrossRef][PubMed]
    [Google Scholar]
  33. Chen IJ, Yuann JM, Chang YM, Lin SY, Zhao J et al. Crystal structure-based exploration of the important role of Arg106 in the RNA-binding domain of human coronavirus OC43 nucleocapsid protein. Biochim Biophys Acta 2013;1834:1054–1062 [CrossRef][PubMed]
    [Google Scholar]
  34. Ma Y, Tong X, Xu X, Li X, Lou Z et al. Structures of the N- and C-terminal domains of MHV-A59 nucleocapsid protein corroborate a conserved RNA-protein binding mechanism in coronavirus. Protein Cell 2010;1:688–697 [CrossRef][PubMed]
    [Google Scholar]
  35. Lin CM, Saif LJ, Marthaler D, Wang Q. Evolution, antigenicity and pathogenicity of global porcine epidemic diarrhea virus strains. Virus Res 2016;226:20–39 [CrossRef][PubMed]
    [Google Scholar]
  36. Yu J, Chai X, Cheng Y, Xing G, Liao A et al. Molecular characteristics of the spike gene of porcine epidemic diarrhoea virus strains in Eastern China in 2016. Virus Res 2018;247:47–54 [CrossRef][PubMed]
    [Google Scholar]
  37. Zhang X, Zhu Y, Zhu X, Shi H, Chen J et al. Identification of a natural recombinant transmissible gastroenteritis virus between Purdue and Miller clusters in China. Emerg Microbes Infect 2017;6:e74 [CrossRef][PubMed]
    [Google Scholar]
  38. Han X, Khan FA, Zhu X, Zhang R, Mustafa R et al. Establishment of an antibody avidity test to differentiate vaccinated cattle from those naturally infected with Mycoplasma bovis. Vet J 2015;203:79–84 [CrossRef][PubMed]
    [Google Scholar]
  39. Shen Z, Ye G, Deng F, Wang G, Cui M et al. Structural basis for the inhibition of host gene expression by porcine epidemic diarrhea virus nsp1. J Virol 2018;92: [CrossRef][PubMed]
    [Google Scholar]
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