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Archives of Virology

, Volume 163, Issue 7, pp 1831–1839 | Cite as

An improved indirect ELISA for specific detection of antibodies against classical swine fever virus based on structurally designed E2 protein expressed in suspension mammalian cells

  • Shengwei Ji
  • Yuzi Luo
  • Tengteng Zhang
  • Lina Shao
  • Xing-Yu Meng
  • Yimin Wang
  • Yao Gao
  • Yongfeng Li
  • Su Li
  • Yuan Sun
  • Xin Jin
  • Hua-Ji Qiu
Original Article
  • 276 Downloads

Abstract

Classical swine fever (CSF), which is caused by classical swine fever virus (CSFV), is a highly contagious disease of pigs. CSFV is genetically and serologically related to bovine viral diarrhea virus (BVDV), a ruminant pestivirus. However, currently available ELISAs based on the full-length E2 protein of CSFV cannot discriminate anti-CSFV from anti-BVDV antibodies. In this study, a truncated CSFV E2 protein (amino acids 690 to 879) covering antigenic domains B/C/D/A (E2B/C/D/A) was designed based on homologous modeling according to the crystal structure of the BVDV E2 protein. The E2B/C/D/A protein was expressed in CHO cells adapted to serum-free suspension culture, and an indirect ELISA (iELISA) was established based on the recombinant protein. No serological cross-reaction was observed for anti-BVDV sera in the iELISA. When testing 282 swine serum samples, the iELISA displayed a high sensitivity (119/127, 93.7%) and specificity (143/155, 92.3%), with an agreement of 92.9% (262/282) and 92.2% (260/282) with virus neutralization test and the IDEXX CSFV blocking ELISA, respectively. Taken together, the newly developed iELISA is highly specific and sensitive and able to differentiate anti-CSFV from anti-BVDV antibodies.

Notes

Acknowledgements

We thank Prof. Paul Becher at the EU Reference Laboratory for CSF, Hannover, Germany, for providing anti-BVDV sera.

Funding

This work was supported by grants from the National Natural Science Foundation of China (No. 31402194 and 31572540) and Harbin Special Fund for Innovation Talents of Science and Technology (2015RAQYJ064).

Compliance with ethical standards

Conflict of interest

All authors declare that they have no competing interests.

Ethical approval

None of the experiments in this study involved human participants.

Supplementary material

705_2018_3809_MOESM1_ESM.docx (42 kb)
Supplementary material 1 (DOCX 41 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute (HVRI)Chinese Academy of Agricultural SciencesHarbinChina
  2. 2.Department of Animal MedicineAgricultural College of Yanbian UniversityYanjiChina

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