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An improved enzyme linked immunosorbent assay for detection of anti-ranavirus antibodies in the serum of the giant toad (Bufo marinus) |
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| Authors: | Z. Zupanovic G. Lopez A. Hyatt B. J. Shiell A. J. Robinson |
| Affiliation: | 1 Australian Animal Health Laboratory (AAHL), PO Bag 24, Geelong, Victoria 3220, Australia 2 Centro de Microbiologia y Biologia Celular, Instituto Venezolano de Investigaciones Cientificas (IVIC), Apartado Postal 21827, Caracas, Venezuela 3 Division of Wildlife and Ecology, Commonwealth Scientific and Industrial Research Organisation, P.O. Box 84, Lyneham, Canberra, 2602, Australia |
| Abstract: | An improved ranavirus antibody ELISA (R Ab ELISA) for the specific detection of anti-ranavirus antibodies in toad sera was developed. Sheep anti-epizootic haematopoietic necrosis virus (EHNV) was used as the antigen-capture antibody. EHNV was used as the antigen and sera from field and challenged toads were used to detect the virus. Rabbit anti-toad IgG and IgM were used to detect bound toad antibody. Pre-absorption of toad sera with a monoclonal antibody, raised against the 50 kDa EHNV protein, improved the specificity of the technique. A blocking ELISA, immunofluorescence and immuno-electron microscopy were used to confirm the validity of the ELISA. The assay has potential use in screening sera from Bufo marinus for the presence of antibodies against ranaviruses and to facilitate understanding of the humoral immunological response in toads during virus infection. |
| Keywords: | Ranavirus antibody ELISA (R Ab ELISA) Bufo marinus Iridovirus Ranavirus |
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