Abstract

A monoclonal antibody-based antigen-capture enzyme-linked immunosorbent assay (ELISA) was developed for the detection of bluetongue virus (BTV) in cell culture lysates and blood samples from sheep. The monoclonal antibody 3E2 and 1C11 specific to BTV VP7 were used as capture antibody and detection antibody, respectively. The assay has detected BTV 1-22 specifically, and had no cross-reactivity with the closely related epizootic hemorrhagic disease virus (EHDV) serotypes 5. The limit of sensitivity of the assay was 9 ng/ml for purified recombinant BTV VP7 and 10^0.5 TCID₅₀/ml for BTV-5. The coefficient of variation (CV) of intra-assay and inter-assay range from 3.45 to 6.10%. The developed antigen-capture ELISA showed good coincident rate (100%) with INGEZIM BTV DAS in 5 serotypes BTV and 8 blood samples from sheep. Therefore, the antigen-capture ELISA may be useful for testing large number of samples in a convenient and short time.

Key words: Bluetongue virus, antigen-capture enzyme-linked immunosorbent assay (ELISA), serotypes, monoclonal antibody.