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.
Artigo
em Inglês
| WPRIM
| ID: wpr-719490
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 ANDMETHODS:
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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Vírus da Doença de Newcastle
/
Ensaio de Imunoadsorção Enzimática
/
Western Blotting
/
Baculoviridae
/
Sensibilidade e Especificidade
/
Discriminação Psicológica
/
Insetos
/
Peso Molecular
/
Doença de Newcastle
/
Nucleoproteínas
Tipo de estudo:
Estudo diagnóstico
Limite:
Animais
Idioma:
Inglês
Revista:
Clinical and Experimental Vaccine Research
Ano de publicação:
2019
Tipo de documento:
Artigo
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