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J Virol Methods ; 198: 12-7, 2014 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-24374124

RESUMO

Avian paramyxovirus 2 (APMV-2) infections are associated with respiratory diseases in poultry worldwide. The hemagglutination inhibition (HI) test is a useful tool for surveillance and monitoring of this virus. In this study, full-length hemagglutinin (HN) gene of APMV-2 was chemically synthesized based on its published sequence, cloned and expressed in Spodoptera frugiperda insect cells using recombinant baculoviruses. The biological, antigenic and immunogenic properties of the expressed protein were evaluated to assess its ability to produce diagnostic reagents for HI testing. Recombinant APMV-2 HN protein showed two distinct bands with molecular masses of 64 and 75kDa, which showed hemagglutination (HA) and neuraminidase activities, respectively. The recombinant HN (rHN) protein extracted from infected cells produced high HA titers (2(13) per 25µL). HA activity of the protein was inhibited by APMV-2 antiserum, although there were weak cross reactions with other APMV serotype antisera. The rHN protein induced high titers of APMV-2-specific antibodies in immunized chickens based on the HI test. These results indicated that recombinant APMV-2 HN protein is a useful alternative to the APMV-2 antigen in HI assays.


Assuntos
Avulavirus/genética , Baculoviridae/genética , Proteína HN/genética , Sequência de Aminoácidos , Animais , Anticorpos Antivirais/imunologia , Antígenos Virais/genética , Antígenos Virais/imunologia , Avulavirus/imunologia , Infecções por Avulavirus/imunologia , Infecções por Avulavirus/prevenção & controle , Infecções por Avulavirus/virologia , Sequência de Bases , Linhagem Celular , Galinhas/imunologia , Galinhas/virologia , Clonagem Molecular/métodos , Reações Cruzadas/imunologia , Proteína HN/imunologia , Testes de Inibição da Hemaglutinação/métodos , Soros Imunes/imunologia , Dados de Sequência Molecular , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/prevenção & controle , Doenças das Aves Domésticas/virologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Células Sf9 , Spodoptera/genética
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