【摘要】 利用Echinococcus granulosus(Eg)成虫表膜抗原抗体夹心ELISA方法检测犬细粒棘球绦虫感染的粪抗原。首先提取E g成虫表膜抗原,制备抗E g成虫表膜抗原的血清,然后用间接ELISA和Western blotting方法检测其抗原抗体反应及免疫原性,用间接免疫组化方法对E g成虫表膜蛋白进行组织定位。利用双抗体夹心ELISA方法检测感染E g的犬粪抗原。ELISA结果表明E g成虫表膜抗原具有良好的抗原性并能产生高效价抗体,抗体不与多头绦虫和泡状带绦虫抗原反应;Western blotting结果表明E g成虫表膜抗原与原头蚴、不成熟E g有共同蛋白成分。免疫组化结果证实虫体表膜蛋白分布在虫体表皮。该抗原抗体系统能检测到感染后13d的犬细粒棘球绦虫。结果表明该E g成虫表膜抗原抗体系统可用于诊断犬细粒棘球绦虫感染。更多还原

【Abstract】 The study was aimed at determining whether antibodies against worm tegument antigens can be used for copro-ELISA to detect Echinococcus granulosus in dogs.In order to produce specific antibodies used for sandwich(copro)-ELISA,we isolated antigens from E.granulosus adult worm tegument.The proteins were used to immunize mice,rabbits and sheep to generate antibodies.ELISA and Western blot analysis were used to identify antigenicity and quality of the serum antibodies.Immunohistochemistry was used to indentify the localization of the proteins in the parasite.Copro-ELISA was used to determine the specificity of the immunodiagnosis for detecting E.granulosus.infection in dogs.Serum titer analysis showed that E.granulosus tegument antigens were antigenic with serum titers reaching to 1:(80 000-320 000) by ELISA.The antibodies recognized antigens from different stages of the parasite.Immunolocalization analysis revealed that the antigens located in the tegument of adult E.granulosus.The antibodies specifically recognized the antigens from E.granulosus with no reaction to the proteins isolated from Multiceps multicepes and T.hydatigena.The results suggest that E.granulosus tegument antigens are a source of antigens for generating antibodies for copro-ELISA to detect the worm antigens in dog feces.更多还原