Figure 1.
Characterization of monoclonal antibody 1F5 by immunofluorescence assay.
Monoclonal antibody 1F5 was used to perform indirect immunofluorescence assay on DF-1 cells infected with DTMUV FX2010 and 293T cells transfected with pCAGGS-E plasmids. A) DF-1 cells infected with DTMUV FX2010, B) control DF-1 cells, C) 293T cells transfected with recombinant plasmid pCAGGS-E, and D) control 293T cells fixed with 4% paraformaldehyde, and then incubated with mAb 1F5 and FITC-conjugated goat anti-mouse IgG, in turn. Cells were mounted with 10 mM p-phenylenediamine (PPD) in glycerol-PBS and observed under a fluorescent microscope.
Figure 2.
Specificity of the b-ELISA to anti-DTMUV serum.
To test the abilities of mAb to bind specifically to domain III of E protein, western blot was conducted with purified fusion protein including both the domain III (12 kDa) of E protein and TF tag protein (52 kDa) (line 1), and purified TF tag protein (52 kDa) (line 2) expressed by pCold plasmids. The mAb 1F5 was able to bind specifically to the 64-kDa fusion protein, but could not bind to purified TF tag protein (Fig 2).
Figure 3.
Specificity of the b-ELISA to anti-DTMUV serum.
Seven antisera against different viruses were investigated. The PI value of anti-DTMUV serum reached a maximum of 69.13%, while the other antisera against H5N1 AIV, H9N2 AIV, NDV, DHV-1, DPV, RV, and JEV ranged from −2.7% to 2.3%.
Table 1.
Comparison of percent inhibition obtained in blocking ELISA and SNT using a serially diluted duck anti-DTMUV serum.
Table 2.
Results of B-ELISA, I-ELISA and SNT with reference sera.
Table 3.
Results of testing field –origin duck sera in blocking ELISA and SNT.