Development of Enzyme-Linked Immunosorbent Assay to Detect Antibodies against Chicken Infectious Anemia Virus

Abstract

standardized and robust indirect enzyme-linked immunosorbent assay (ELISA) to detect antibodies against chicken infectious anemia virus (CIAV) was developed. Thai CIAV was isolated and used for the entirety of
this study. For virus cultivation, a cell density of MDCC-MSB1 cells at 3 x 106 cells/ml and a fetal bovine serum concentration at 10 percent were the most appropriate for Thai CIAV. Then, crude virus of CIAV was prepared using ultracentrifugation technique. To develop the ELISA test, optimal dilution of goat anti-chicken IgG was 1 : 200 and antigen concentration in the ELISA was 1.00 μg/ml, as determined by checkerboard titration. In the present study, a comparison was made between commercial and in-house ELISA for the detection of antibodies to CIAV. Relative sensitivity, specificity and accuracy of in-house ELISA were 93%, 78% and 86%, respectively. Agreement between commercial and in-house ELISA was substantial (Kappa value = 0.71). Consequently, the in-house ELISA was as good as the commercial ELISA in screening chicken serum samples for antibodies against CIAV.