Development of a dot-enzyme linked immunosorbent assay for detection of bovine antibody against Clostridium perfringens type D.

A dot-enzyme linked immunosorbent assay(dot-ELISA) for detection of cattle's antibody against Clostridium perfringens type D using purified antigen was developed.The optimal conditions for the dot-ELISA assay were that antigen concentration,serum dilution and dilution of the HRP-labeled rabbit anti-bovine IgG were 1.95 μg/mL,1∶320,1∶2 000,respectively and blocking agent was 30 mL/L skimmed milk at 37 ℃ for 30 min.Positive rate of 30 bovine serum samples in dot-ELISA using the purified antigen was 16.7% higher than that using the crude antigen.Sensitivity of the dot-ELISA was 43 times that of AGID.Positive rate of 100 bovine serum samples collected from Yanbian Prefecture was 59.0% by dot-ELISA.Specificity,reproducibility and comparison tests showed that dot-ELISA was specific,sensitive and reproducible.