Virologica Sinica

, Volume 32, Issue 4, pp 307–316 | Cite as

Evaluation of purified recombinant spike fragments for assessment of the presence of serum neutralizing antibodies against a variant strain of porcine epidemic diarrhea virus

  • Jianwei Hao
  • Yun Zhang
  • Shengkun Fang
  • Zhifen Wen
  • Xiangbin Zhang
  • Chunyi Xue
  • Yongchang Cao
Research Article

Abstract

Since 2010, variant strains of porcine epidemic diarrhea virus (PEDV) have caused disasters in the pork industry. The spike (S) protein, as the major immunity-eliciting antigen, has previously been used for serological testing and has been found to correlate significantly with the results of the serum neutralization (SN) test. However, further evaluation of this method is needed as new epidemic strains of PEDV emerge. Hence, the main objective of this study was to assess sow sera and determine the correlation between enzyme-linked immunosorbent assay (ELISA) results (involving a newly isolated GDS01 virus-based ELISA and ELISAs based on seven recombinant fragments comprising overlapping S1 and partial S2 sequences) and SN titers. Furthermore, we determined the reliability of the ELISAs based on receiver operating characteristics (ROC) curve analyses. For the most promising ELISA, i.e., the SP4 ELISA, the correlation coefficient (r) and the area under curve (AUC) were determined to be 0.6113 and 0.8538, respectively. In addition, we analyzed the homology of the SP4 sequences obtained from different strains (including vaccine strains) and found that various strains showed a high degree of homology in this region. Thus, we conclude that SP4 is a promising serological testing protein for use in the field.

Keywords

Porcine epidemic diarrhea virus (PEDV) spike protein enzyme-linked immunosorbent assay serum neutralization test 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Supplementary material

12250_2017_3969_MOESM1_ESM.pdf (1.1 mb)
Evaluation of purified recombinant spike fragments for assessment of the presence of serum neutralizing antibodies against a variant strain of porcine epidemic diarrhea virus

References

  1. Chang SH, Bae JL, Kang TJ, Kim J, Chung GH, Lim CW, Laude H, Yang MS, Jiang YS. 2002. Identification of the Epitope Region Capable of Inducing Neutralizing Antibodies against the Porcine Epidemic Diarrhea Virus. Mol Cells, 14: 295–299.PubMedGoogle Scholar
  2. Chen F, Pan Y, Zhang X, Tian X, Wang D, Zhou Q, Song Y, Bi Y. 2012. Complete genome sequence of a variant porcine epidemic diarrhea virus strain isolated in China. J Virol, 86: 12448.CrossRefPubMedPubMedCentralGoogle Scholar
  3. Chen JF, Liu XZ, Shi D, Shi HY, Zhang X, Li CL, Chi YB, Feng L. 2013. Detection and molecular diversity of spike gene of porcine epidemic virus in China. Viruses, 5: 2601–2613.CrossRefPubMedPubMedCentralGoogle Scholar
  4. Duarte M, Tobler K, Bridgen A, Rasschaert D, Ackermann M, Laude H. 1994. Sequence analysis of the porcine epidemic diarrhea virus genome between the nucleocapsid and spike protein genes reveals a polymorphic ORF. Virology, 198: 466–476.CrossRefPubMedGoogle Scholar
  5. Egberink HF, Ederveen J, Callebaut P, Horzinek MC. 1988. Characterization of the structural proteins of porcine epizootic diarrhea virus, strain CV777. Am J Vet Res, 49: 1320–1324.PubMedGoogle Scholar
  6. Fan H, Zhang J, Ye Y, Tong T, Xie K, Liao M. 2012. Complete genome sequence of a novel porcine epidemic diarrhea virus in south China. J Virol, 86: 10248–10249.CrossRefPubMedPubMedCentralGoogle Scholar
  7. Ge JW, Liu DQ, Li YJ. 2012. Construction of recombinant lactobacilli expressing the core neutralizing epitope (COE) of porcine epidemic diarrhea virus and a fusion protein consisting of COE and Escherichia coli heat-labile enterotoxin B, and comparison of the immune responses by orogastric immunization. Can J Microbiol, 58: 1258–1267.CrossRefPubMedGoogle Scholar
  8. Gibertoni AM, Montassier MF, Sena JA, Givisiez PE, Furuyama CR, Montassier HJ. 2005. Development and application of a Saccharomyces cerevisiae-expressed nucleocapsid proteinbased enzyme-linked immunosorbent assay for detection antibodies against infectious bronchitis virus. J Clin Microbiol, 43: 1982–1984.CrossRefPubMedPubMedCentralGoogle Scholar
  9. Hao JW, Xue CY, He LL, Wang Y, Cao YC. 2014. Bioinformatics insight into the spike glycoprotein gene of field porcine epidemic diarrhea strains during 2011-2013 in Guangdong, China. Virus genes. Virus Genes, 49: 58–67.CrossRefPubMedGoogle Scholar
  10. Hofmann M, Wyler R. 1990. Enzyme-linked immunosorbent assay for the detection of porcine epidemic diarrhea coronavirus antibodies in swine sera. Vet Microbiol, 21: 263–273.CrossRefPubMedGoogle Scholar
  11. Huang YC, Ho CC, Lin BJ, Lai CH, Liaw YP. 2010. Optimal cutoff value of high-density lipoprotein cholesterol for predicting coronary artery disease in Taiwanese population. Nutr Res, 30: 21–26.CrossRefPubMedGoogle Scholar
  12. Huang YW, W Dickerman, Pineyro P, Li L, Fang L, Kiehne R, Opriessnig T, Meng XJ. 2013. Origin, Evolution, and Genotyping of Emergent Porcine Epidemic Diarrhea Virus Strains in the United States. mBio. doi:10.1128/mBio.00737–13.Google Scholar
  13. Ignjatovic J, Sapats S. 2005. Identification of previously unknown antigenic epitopes on the S and N proteins of avian infectious bronchitis virus. Arch Virol, 150: 1813–1831.CrossRefPubMedGoogle Scholar
  14. Koch G, Hartog L, Kant A, van Roozelaar DJ. 1990. Antigenic domains on the peplomer protein of avian infectious bronchitis virus: correlation with biological functions. J Gen Virol, 71: 1929–1935.CrossRefPubMedGoogle Scholar
  15. Luo Y, Zhang J, Deng X, Ye Y, Liao M, Fan H. 2012. Complete genome sequence of a highly prevalent isolate of porcine epidemic diarrhea virus in south China. J Virol, 86: 9551.CrossRefPubMedPubMedCentralGoogle Scholar
  16. Niesters HG, Lenstra JA, Spaan WJ, Zijderveld AJ, Bleumink-Pluym NM, Hong F, van Scharrenburg G.J, Horzinek MC, van der Zeijst BA 1986. The peplomer protein sequence of the M41 strain of coronavirus IBV and its comparison with Beaudette strains. Virus Res, 5: 253–263.CrossRefPubMedGoogle Scholar
  17. Oh JS, Song DS, Yang JS, Song JY, Moon HJ, Kim TY, Park BK. 2005. Comparison of an enzyme-linked immunosorbent assay with serum neutralization test for serodiagnosis of porcine epidemic diarrhea virus infection. J Vet Sci, 6: 349–352.PubMedGoogle Scholar
  18. Oszvald M, Kang TJ, Tomoskozi S, Tamas C, Tama L, Kim TG, Yang MS. 2007. Expression of a synthetic neutralizing of porcine epidemic diarrhea virus fused with synthetic B subunit of Escherichia coli heat labile enterotoxin in rice endosperm. Mol Biotechnol, 35: 215–223.CrossRefPubMedGoogle Scholar
  19. Park SJ, Moon HJ, Yang JS, Lee CS, Song DS, Kang BK, Park BK. 2007. Sequence analysis of the partial spike glycoprotein gene of porcine epidemic diarrhea viruses isolated in Korea. Virus Genes, 35: 321–332.CrossRefPubMedGoogle Scholar
  20. Song D, Park B. 2012. Porcine epidemic diarrhoea virus: a comprehensive review of molecular epidemiology, diagnosis, and vaccines. Virus Genes, 44: 167–175.CrossRefPubMedGoogle Scholar
  21. Spaan W, Cavanagh D. 1988. Coronavirus: structure and genome expression. J Gen Virol, 69: 29–52.CrossRefGoogle Scholar
  22. Sucheeva Junnu, Preeda Lertwatchrasarakul, Siriluk Jala, Sakuna Phattanakunana, Pattra Moonjit, Thaweesak Songserm. 2013. Developing an indirect ELISA based o recombinant hexon protein for serological detection of inclusion body hepatitis in chickens. J Vet Med Sci, 76: 289–293.Google Scholar
  23. Sun D, Feng L, Shi H, Chen J, Cui X, Chen H, Liu S, Tong Y, Wang Y, Tong G. 2008. Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein. Vet Microbiol, 131: 73–81.CrossRefPubMedGoogle Scholar
  24. Tian YF, Yu ZJ, Cheng KH, Liu YX, Huang J, Xin Y, Li YH, Fan ST, Wang TC, Huang G, Feng N, Yang ZG, Yang ST, Gao YW, Xia XZ. 2013. Molecular characterization and phylogenetic analysis of new variants of the porcine epidemic diarrhea virus in Gansu China in 2012. Viruses, 5: 1991–2004.CrossRefPubMedPubMedCentralGoogle Scholar
  25. Wei ZY, Lu WH, Li ZL, Mo JY, Zeng XD, Zeng ZL, Sun BL, Chen F, Xie QM, Bee YZ, Ma JY. 2012. Complete genome sequence of novel porcine epidemic diahrrhea virus stain GD-1 in China. J Virol, 86: 13824–13825.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Wuhan Institute of Virology, CAS and Springer Nature Singapore Pte Ltd. 2017

Authors and Affiliations

  1. 1.State Key Laboratory of Biocontrol, School of Life SciencesSun Yat-sen UniversityGuangzhouChina
  2. 2.Fraunhofer-Institut für Angewandte Informationstechnik FITSankt AugustinGermany

Personalised recommendations