Oral administration of inactivated porcine epidemic diarrhea virus activate DCs in porcine Peyer’s patches

Yuan, Chen; Zhang, En; Huang, Lulu; Wang, Jialu; Yang, Qian

The PEDV-specific IgA in feces and PEDV-specific IgG in serum were detected by ELISA. ELISA plates were coated 2 μg purified PEDV/well at 4°C overnight. After antigen removal, ELISA plates were blocked with 3% bovine serum albumin (BSA) in PBS which contains 0.05% Tween (PBST) for 2 h at 37°C. Then, 100-fold dilutions of serum samples or 2-fold dilutions of lavage fluid from pig were added to the plates and incubated at 37°C for 2 h. Washed with PBST and added 100 μl of HRP-conjugated goat anti-pig IgA/IgG antibody at 1:2000 dilution and incubated at 37°C for 1 h. Plates were washed 5 times and incubated with 3, 3′, 5, 5′-tetramethylbenzidine (TMB) for 15 min. Then, the reaction was stopped with sulfuric acid (2 M). Optical densities at 450 nm were measured using an enzyme-linked immunosorbent assay (ELISA) plate reader.