Standardization of an Indirect Enzyme Linked Immuno Sorbent Assay for Measuring Antibodies of Infectious Bursal Disease Virus

An attempt was made to standardize an in-house immuno-enzyme assay for measuring antibodies of infectious bursal disease virus (IBDV) in chicken. The test was performed after coating plates with ELISA antigens prepared by two methods. Antigen “A” was prepared from infectious bursal disease virus infected chicken embryo fibroblasts culture by concentration with dialysis against PEG-6000 while antigen “B” was prepared reconstituting a live infectious bursal disease virus vaccine. The optimum dilution of antigen “A” and “B” was found to be 1:300 and 1:600 respectively. Both the antigens produced acceptable and comparable results but antigen “B” is conventional due to ease of preparation and to avoid a time consuming and costly procedure of cell culture. The rabbit anti-chicken immunoglobulin-G conjugated to horseradish peroxidase was used at a dilution of 1:2000. The assay was evaluated by testing chicken serum samples of different age groups (1-day-old broiler breeder and broiler chicks, 13 weeks old vaccinated layer breeder birds and 30 week old vaccinated broiler breeder birds). The efficiency of the standardized ELISA was compared with a commercially available ELISA kit. The results indicated that in-house developed ELISA was equally as sensitive and specific as commercially available kit in detection of antibodies against IBDV.