Detection of major capsid protein of infectious myonecrosis virus in shrimps using monoclonal antibodies.
AUTOR(ES)
SEIBERT, C. H.
FONTE
Journal of Virological Methods
DATA DE PUBLICAÇÃO
2011
RESUMO
Infectious myonecrosis virus (IMNV) has been causing a progressive disease in farm-reared shrimps in Brazil and Indonesia. Immunodiagnosticmethods for IMNVdetection, although reliable, are not employed currently becausemonoclonal antibodies (MAbs) against this virus are not available. In this study, a fragment of the IMNVmajor capsid protein gene, comprising amino acids 300?527 (IMNV300?527),was cloned and expressed in Escherichia coli. The nucleotide sequence of the recombinant IMNV300?527 fragment displayed a high degree of identity to the major capsid protein of IMNV isolates from Brazil (99%) and Indonesia (98%). Ten MAbs were generated against the expressed fragment, and eight of these, mostly IgG2a or IgG2b,were able to bind to IMNVin tissue extracts fromshrimps infected naturally in immunodotblot assays. Six of these MAbs recognized a ?100 kDa protein in a Western-blot, which is the predicted mass of IMNV major capsid protein, and also bound to viral inclusions present in muscle ?broses and in coagulativemyonecrosis, as demonstrated by immunohistochemistry. Among all thoseMAbs created, four did not cross-react with non-infected shrimp tissues; this observation supports their applicability as a sensitive and speci?c immunodiagnosis of IMNV infection in shrimps.
ASSUNTO(S)
vírus da mionecrose infecciosa camarão proteína recombinante anticorpo monoclonal
ACESSO AO ARTIGO
http://www.alice.cnptia.embrapa.br/handle/doc/870999Documentos Relacionados
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