Peng Z, Simons F E, Becker A B
Department of Paediatrics and Children's Health, University of Manitoba, Winnipeg, Canada.
Am J Vet Res. 1993 Feb;54(2):239-43.
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.
我们使用多克隆兔抗犬IgE抗体和单克隆小鼠抗犬IgE抗体，开发了用于测量犬血清中豚草特异性IgE的酶联免疫吸附测定法（ELISA）。在该ELISA中，微量滴定板用豚草提取物包被，然后依次与犬血清、纯化的单克隆或多克隆抗犬IgE以及与小鼠IgG或兔IgG结合的山羊抗体孵育。通过该ELISA测量了60只豚草过敏犬的血清样本和10只对照犬的血清中豚草特异性IgE值。对这些血清进行了被动皮肤过敏反应（PCA）试验，以将结果与ELISA的结果进行比较。使用多克隆抗体的测定法之间的平均变异系数为0.20±0.10，使用单克隆抗体的测定法之间的平均变异系数为0.17±0.10。使用多克隆抗体的ELISA的灵敏度为0.6 U/ml，使用单克隆抗体的ELISA的灵敏度为2.5 U/ml。通过这两种ELISA测量的血清豚草特异性IgE值与PCA滴度高度相关（P<0.0000），但使用多克隆抗体的ELISA与PCA滴度的相关性（r = 0.84）高于使用单克隆抗体的ELISA（r = 0.59）。通过这两种ELISA和PCA试验测量的豚草特异性IgE几何平均值，在过敏犬中显著高于（P<0.0000）对照犬。这两种ELISA在测量犬血清中豚草特异性IgE方面具有特异性、敏感性和可重复性。
