Sundquist B, Bech-Nielsen S, Zakrisson G
National Veterinary Institute, Uppsala, Sweden.
Vet Parasitol. 1989 Oct;33(3-4):309-18. doi: 10.1016/0304-4017(89)90140-4.
A study was conducted to determine if the purification of Parafilaria bovicola antigens can increase the specificity of serodiagnosis of parafilariasis in enzyme-linked immunosorbent assay. Antigens released from adult worms of P. bovicola were separated by chromatofocusing on a polybuffer exchanger of the pH range 7.3-4.0 Polypeptide analysis by sodium dodecyl sulphate polyacrylamide gel electrophoresis showed the presence of four major polypeptides with MWs of 41, 36, 24 and 20 kDa. Additional biochemical characterization identified the 24- and 20-kDa polypeptides as hydrophobic glycoproteins. The chromatofocusing purification procedures were also applied for separation of a whole-worm extract. Again, the 41- and 36-kDa antigens were identified in separate peak fractions. Using ELISA, it was shown that the 41- and 35-kDa antigens were recognized by bovine antibodies specific for P. bovicola, but not by other sera collected from cattle infected by Onchocerca gutturosa, Onchocerca lienalis, Ostertagia ostertagi and Dictyocaulus viviparus. The serological evaluation strongly suggests that the 41- and 36-kDa antigens are P. bovicola specific.
开展了一项研究，以确定牛副丝虫抗原的纯化是否能提高酶联免疫吸附测定中牛副丝虫病血清诊断的特异性。通过在pH范围为7.3 - 4.0的多缓冲交换剂上进行色谱聚焦，分离牛副丝虫成虫释放的抗原。十二烷基硫酸钠聚丙烯酰胺凝胶电泳进行的多肽分析显示存在四种主要多肽，分子量分别为41、36、24和20 kDa。进一步的生化特性鉴定表明，24 kDa和20 kDa的多肽为疏水糖蛋白。色谱聚焦纯化程序也用于分离全虫提取物。同样，在单独的峰级分中鉴定出了41 kDa和36 kDa的抗原。使用酶联免疫吸附测定表明，41 kDa和35 kDa的抗原能被牛副丝虫特异性牛抗体识别，但不能被从感染了喉瘤盘尾丝虫、线性盘尾丝虫、奥斯特他线虫和胎生网尾线虫的牛采集的其他血清识别。血清学评估强烈表明，41 kDa和36 kDa的抗原是牛副丝虫特异性的。
