Detection of infectious myonecrosis virus in penaeid shrimps using immunoassays: usefulness of monoclonal antibodies directed to the viral major capsid protein.
Despite the economic impact of the infectious myonecrosis virus (IMNV) on shrimp farms in several countries, no method for immunological detection is currently available. With the aim of developing immunodiagnostic methods for IMNV detection in infected shrimps, a recombinant fragment of the IMNV major capsid protein gene encoding amino acids 105-297 (rIMNV₁₀₅₋₂₉₇ was heterologously expressed in Escherichia coli and used to immunize Balb/c mice, generating monoclonal antibodies (MAbs). Six hybridomas were obtained, and four of these recognized the presence of IMNV in tissue homogenates from naturally infected shrimps by immunodot blot assay. Among these MAbs, three were able to detect a ~100-kDa protein, which corresponds to the predicted mass of the IMNV major capsid protein, as well as viral inclusion bodies in muscle fibroses by western blot and immunohistochemistry. Two MAbs showed high specificity and sensitivity, showing no cross-reaction with healthy shrimp tissues in any assays, indicating their usefulness for IMNV detection.
Borsa
Mariana
M
Departamento de Microbiologia, Imunologia e Parasitologia, Universidade Federal de Santa Catarina, UFSC, Florianópolis, Brazil.
Seibert
Caroline H
CH
Rosa
Rafael D
RD
Stoco
Patrícia H
PH
Cargnin-Ferreira
Eduardo
E
Pereira
Alitiene M L
AM
Grisard
Edmundo C
EC
Zanetti
Carlos R
CR
Pinto
Aguinaldo R
AR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D036022:Capsid Proteins / Q000276:immunology*; D003001:Cloning, Molecular; D007118:Immunoassay / Q000662:veterinary*; D007150:Immunohistochemistry; D034582:Integumentary System / Q000821:virology; D051379:Mice; D008807:Mice, Inbred BALB C; D033561:Penaeidae / Q000821:virology*; D012367:RNA, Viral; D019196:Totiviridae / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D036022:Capsid Proteins; D012367:RNA, Viral