Figure 3

Reactivity of N3T and N7T antigens with (a) neuraminidase-positive AIV serum and (b) hemagglutinin-positive AIV serum. The ELISA plates were coated with N3T and N7T antigens at 60 μg/ml and 50 μg/ml, respectively. The coated plates were reacted with serum samples at single 1:100 dilution and incubated for 30 min at room temperature, washed three times with PBST (PBS with 0.05% Tween-20), followed by 30 min incubation with goat anti-chicken IgG HRP-conjugated secondary antibody. The reaction was developed with ABTS (2, 2'-Azinobis [3-thylbenzothiazoline-6-sulfonic acid]-diammonium salt) substrate, stopped after 15 minutes with 1% SDS, and the plates were read in a spectrophotometer using the filter at 405 nm.