Detection of major capsid protein of infectious myonecrosis virus in shrimps using monoclonal antibodies

作者：Seibert Caroline H; Borsa Mariana; Rosa Rafael D; Cargnin Ferreira Eduardo; Pereira Alitiene M L; Grisard Edmundo C; Zanetti Carlos R; Pinto Aguinaldo R^*

来源：Journal of Virological Methods, 2010, 169(1): 169-175.

DOI：10.1016/jjviromet.2010.07.020

• Infectious myonecrosis virus (IMNV)
• Recombinant protein
• Monoclonal antibody
• lmmunodiagnostic methods
• Penaeid shrimp

Infectious myonecrosis virus (IMNV) has been causing a progressive disease in farm-reared shrimps in Brazil and Indonesia. Immunodiagnostic methods for IMNV detection, although reliable, are not employed currently because monoclonal antibodies (MAbs) against this virus are not available. In this study, a fragment of the IMNV major capsid protein gene, comprising amino acids 300-527 (IMNV(300-527)), 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 IgG(2a) or IgG(2b), were able to bind to IMNV in tissue extracts from shrimps infected naturally in immunodotblot assays. Six of these MAbs recognized a similar to 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 fibroses and in coagulative myonecrosis, as demonstrated by immunohistochemistry. Among all those MAbs created, four did not cross-react with non-infected shrimp tissues; this observation supports their applicability as a sensitive and specific immunodiagnosis of IMNV infection in shrimps.

• 出版日期2010-10