Measurement of ragweed-specific IgE in canine serum by use of enzyme-linked immunosorbent assays, containing polyclonal and monoclonal antibodies
1993
Using polyclonal rabbit and monoclonal mouse anti-dog IgE antibodies, we developed ELISA for measurement of ragweed-specific IgE in canine serum. In the ELISA, microtitration plates were coated with ragweed extract and sequentially incubated with canine serum, purified monoclonal or polyclonal anti-dog IgE, and conjugated goat antibody to mouse IgG or rabbit IgG. Serum ragweed-specific IgE values were measured by the 2 ELISA in serum samples from 60 ragweed-allergic dogs and in serum from 10 control dogs. Passive cutaneous anaphylaxis (PCA) tests were performed on these sera to compare results with those of the ELISA, Mean coefficient of variation between assays was 0.20 +/- 0.10 for the assay using the polyclonal antibody and was 0.17 +/- 0.10 for that using monoclonal antibody. Sensitivity was 0.6 U/ml for the ELISA, using polyclonal antibody, and 2.5 U/ml for the ELISA, using monoclonal antibody. Serum ragweed-specific IgE values measured by the 2 ELISA strongly correlated with PCA titers (P < 0.0000), but the ELISA using polyclonal antibody had higher correlation with PCA titer (r = 0.84) than the ELISA using monoclonal antibody (r = 0.59). The geometric mean ragweed-specific IgE value measured by the 2 ELISA and by PCA testing, was significantly higher (P < 0.0000) in allergic dogs than in control dogs. The 2 ELISA were specific, sensitive, and reproducible for measurement of ragweed-specific IgE in canine serum.
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