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Following incubation 100 μl of mice sera samples, positive and negative control sera diluted 1 200 were added into the plates in duplicate wells and incubated at 37 °C for 1 h.
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For the isotype-specific ELISA, after incubation with the mouse sera samples (1 : 200 dilution), the plates were incubated with goat anti-mouse IgG1 or IgG2a (BD Pharmingen, USA).
Two peptides, one each from WNV (named WE147) and JEV (named JE40) E-protein, showed virus-specific and strong reactivity to the immune mice sera and human clinical samples.
After this blocking step, the wells were washed five times with PBS-T and then 100 μL/well of sera samples from mice diluted 1 100 in PBS-T were added in duplicate.
Sera samples from the immunised mice were used in immunoassays (ELISA) for detection of specific IgG1, IgG2a and IgG3 antibodies.
Dot blotting of EC mice sera with an anti-mice AFP antibody showed a positive reaction in all EC samples (B), but not in the control 35 Ds sera (C in B).
Dot blotting of control and EC mice sera with an anti-mice AFP antibody sc-81088) shows a positive reaction in the EC samples, but not in the control sera (Fig. 4D).
Sera samples collected from three groups of mice during the experiment period (0 63 days), in addition to negative control samples were tested using semi-quantitative RBPT as described by OIE [ 35] with some modifications.
Measurement of IL-27levels in sera samples was conducted by using a Quantikine kit for mouse IL-27 p28 (R&D Systems).
Table 2 shows the recovery of ricin from spiked mouse sera and feces samples.
(d) Spearman's correlation of #162 binding signals and gp41-Min titer of immunized mice sera.
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