Expression and serological application of recombinant epitope-repeat protein carrying an immunodominant epitope of Newcastle disease virus nucleoprotein
Clinical and Experimental Vaccine Research
; : 27-34, 2019.
Article
in En
| WPRIM
| ID: wpr-719490
Responsible library:
WPRO
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.
Key words
Full text:
1
Index:
WPRIM
Main subject:
Newcastle disease virus
/
Enzyme-Linked Immunosorbent Assay
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Blotting, Western
/
Baculoviridae
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Sensitivity and Specificity
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Discrimination, Psychological
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Insecta
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Molecular Weight
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Newcastle Disease
/
Nucleoproteins
Type of study:
Diagnostic_studies
Limits:
Animals
Language:
En
Journal:
Clinical and Experimental Vaccine Research
Year:
2019
Type:
Article