DAO 37:99-106 (1999) - doi:10.3354/dao037099
Production and use of antibodies for the detection of Taura syndrome virus in penaeid shrimp
B. T. Poulos1,*, R. Kibler2, D. Bradley-Dunlop1, L. L. Mohney1, D. V. Lightner1
ABSTRACT: Monoclonal (MAb) and polyclonal (PAb) antibodies were produced against the penaeid shrimp virus, Taura syndrome virus (TSV), isolated from naturally infected Penaeus vannamei from farms in Ecuador, Hawaii and Texas. The PAbs produced in both chickens and mice were capable of detecting TSV in the hemolymph of shrimp during the acute phase of infection by an immunoblot assay. The MAbs were produced using BALB/cByJ mouse spleen cells fused with non-immunoglobulin-secreting SP2/0-Ag-14 mouse myeloma cells. Three MAbs of different immunoglobulin isotypes (IgGγ 1κ, IgG 2bκ and IgG 3κ) were compared in an immunoblot assay to determine their reactivity to hemolymph from TSV-infected shrimp during the acute and chronic phases of infection and their cross-reactivity, if any, with other shrimp viruses. Western blots of purified TSV were used to compare the specificities of the 3 MAbs for the structural proteins of the virus. The MAbs were used to monitor a laboratory-induced TSV infection in juvenile P. vannamei, and they were capable of detecting the virus in the hemolymph of both acute and chronic phase samples. The results with chronic phase hemolymph samples were variable and indicated the need to develop an immunoassay in which the virus in a sample is captured by one antibody and then detected with a second antibody. The availability of MAbs with different specificities for TSV viral proteins and the ability to produce PAbs in chickens will make it possible to develop a sensitive capture assay for rapid detection of the virus in field situations.
KEY WORDS: TSV · Taura syndrome virus · Monoclonal antibodies · Immunodetection
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