Clin Exp Vaccine Res.  2019 Jan;8(1):27-34. 10.7774/cevr.2019.8.1.27.

Expression and serological application of recombinant epitope-repeat protein carrying an immunodominant epitope of Newcastle disease virus nucleoprotein

Affiliations
  • 1Department of Veterinary Microbiology, College of Veterinary and Animal Sciences, Parbhani, India.
  • 2Avian Disease Research Division, Animal and Plant Quarantine Agency, Gimcheon, Korea.
  • 3Veterinary Drugs and Biologics Division, Animal and Plant Quarantine Agency, Gimcheon, Korea.
  • 4Planning and Coordination Division, Animal and Plant Quarantine Agency, Gimcheon, Korea. kchoi0608@korea.kr

Abstract

PURPOSE
The aim of the present study was to develop a serodiagnostic test for differentiation infected from vaccinated animal (DIVA) strategy accompanying the marker vaccine lacking an immunodominant epitope (IDE) of nucleoprotein of Newcastle disease virus (NDV).
MATERIALS AND METHODS
Recombinant epitope-repeat protein (rERP) gene encoding eight repeats of the IDE sequence (ETQFLDLMRAVANSMR) by tetra-glycine linker was synthesized. Recombinant baculovirus carrying the rERP gene was generated to express the rERP in insect cells. Specificity and sensitivity of an indirect enzyme-linked immunosorbent assay (ELISA) employing the rERP was evaluated.
RESULTS
The rERP with molecular weight of 20 kDa was successfully expressed by the recombinant baculovirus in an insect-baculovirus system. The rERP was antigenically functional as demonstrated by Western blotting. An indirect ELISA employing the rERP was developed and its specificity and sensitivity was determined. The ELISA test allowed discrimination of NDV infected sera from epitope deletion virus vaccinated sera.
CONCLUSION
The preliminary results represent rERP ELISA as a promising DIVA diagnostic tool.

Keyword

Newcastle disease virus; Nucleoprotein; Epitope-repeat protein; Enzyme-linked immunosorbent assay; Differentiation infected from vaccinated animal

MeSH Terms

Animals
Baculoviridae
Blotting, Western
Enzyme-Linked Immunosorbent Assay
Insects
Molecular Weight
Newcastle disease virus*
Newcastle Disease*
Nucleoproteins
Sensitivity and Specificity
Nucleoproteins
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