An improved enzyme linked immunosorbent assay for detection of anti-Ranavirus antibodies in the serum of the giant toad (Bufo marinus)

An improved ranavirus antibody ELISA (R Ab ELISA) for the specific detectio n of anti-ranavirus antibodies in toad sera was developed. Sheep anti-epizo otic haematopoietic necrosis virus (EHNV) was used as the antigen-capture a ntibody. EHNV was used as the antigen and sera from field and challenged to ads were used to detect the virus. Rabbit anti-toad IgG and IgM were used t o 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 immune-electron microscopy were used to confirm the validity of the ELISA, The assay has p otential use in screening sera from Bufo marinus for the presence of antibo dies against ranaviruses and to facilitate understanding of the humoral imm unological response in toads during virus infection. (C) 1998 Elsevier Scie nce Ltd. Ail rights reserved.