de Graaf D C, Berghen P, Hilderson H, Claerebout E, Vercruysse J
Department of Parasitology, Faculty of Veterinary Medicine, University of Gent, Belgium.
Int J Parasitol. 1994 Aug;24(5):681-8. doi: 10.1016/0020-7519(94)90121-x.
Ostertagia ostertagi adult worm extracts were analysed by Western blotting using sera from calves experimentally infected with O. ostertagi and Cooperia oncophora. Strong differences in antigen recognition were noticed, even between animals from the same group. Two Ostertagia specific antigens with apparent molecular mass of 19.7 (OoA19.7) and 20.7 kDa (OoA20.7) were identified. One of them (OoA19.7) was purified by three subsequent chromatographic steps, i.e. gelfiltration, ion-exchange and reversed phased chromatography. It was demonstrated that this antigen does not show any cross-reactions with heterologous sera from C. oncophora, Dictyocaulus viviparus, Nematodirus helvetianus and Fasciola hepatica-infected animals. It was found that only IgG1 antibodies reacted against OoA19.7. The application of this antigen in an ELISA resulted in a highly species-specific test when compared to crude worm extracts. However, strong individual differences in anti-OoA19.7 response could be noticed between calves which received the same number of O. ostertagi larvae. These individual differences can hinder the application of the ELISA as a diagnostic tool, since the anti-OoA19.7 response does not seem to reflect the level of exposure to L3 larvae. This was supported by the absence of a clear infection-dose-related effect. It was shown that the anti-OoA19.7 response started from week 6 to 8, and reached its highest level at week 15.
利用感染奥氏奥斯特线虫(Ostertagia ostertagi)和牛库珀线虫(Cooperia oncophora)的犊牛血清,通过蛋白质免疫印迹法分析奥氏奥斯特线虫成虫提取物。即使是同一组动物之间,抗原识别也存在显著差异。鉴定出两种奥氏奥斯特线虫特异性抗原,表观分子量分别为19.7 kDa(OoA19.7)和20.7 kDa(OoA20.7)。其中一种(OoA19.7)通过连续三步色谱法纯化,即凝胶过滤、离子交换和反相色谱法。结果表明,该抗原与感染牛库珀线虫、胎生网尾线虫(Dictyocaulus viviparus)、瑞士细颈线虫(Nematodirus helvetianus)和肝片吸虫(Fasciola hepatica)动物的异源血清无交叉反应。发现只有IgG1抗体与OoA19.7发生反应。与粗制虫体提取物相比,该抗原用于酶联免疫吸附测定(ELISA)时具有高度的种特异性。然而,在接种相同数量奥氏奥斯特线虫幼虫的犊牛之间,抗OoA19.7反应存在明显的个体差异。这些个体差异可能会阻碍ELISA作为诊断工具的应用,因为抗OoA19.7反应似乎不能反映对L3幼虫的暴露水平。这一点得到了缺乏明确的感染剂量相关效应的支持。结果表明,抗OoA19.7反应从第6周开始,至第8周出现,并在第15周达到最高水平。