粪抗原检测法诊断犬细粒棘球绦虫感染的研究 Ⅰ.用细粒棘球绦虫成虫虫体抗原—抗体系统检测粪抗原方法的建立

Diagnosis of Echinococcus granulosus Infection in Dogs by Using Coproantigen Detection I. Establishment of Methods by Using E.granulosus Adult Sometic Antigen-Antibody System for Coproantigen Detection in Dogs

用细粒棘球绦虫成虫虫体抗原及其免疫家兔血清ＩｇＧ建立了用于检测家犬粪抗原诊断细粒棘球绦虫成虫感染的双抗体夹心ＥＬＩＳＡ方法。对用离子交换层析、ＳＰＡ亲和层析、抗兔ＩｇＧ抗体亲和层析和细粒棘球绦虫成虫虫体抗原亲和层析等四种方法纯化的抗体进行了比较，以抗原亲和层析法纯化的抗体免疫活性最高，用双抗体夹心ＥＬＩＳＡ检测抗原的终点在标本缓冲液中为２．５ｎｇ／ｍｌ，在健康家犬粪便悬液的上清中为５ｎｇ／ｍｌ。检测１１只人工感染细粒棘球绦虫成虫的家犬粪便标本，粪抗原阳性率达１００％。

The establishment of method of double antibody sandwich ELISA for diagnosis of E.granulosus infection in dogs by using the adult somatic antigen-antibody system was reported. The methods of ion exchange chromatography, SPA affinity chromatography, anti-rabbit IgG antibody affinity chromatography and antigen affinity chromatography were used in antibody purification. The best immunoactivities were obtained in the antibodies purified by antigen affinity chromatography. The endpoint of antigen detection by double antibody-sandwich ELISA were 2.5ng/ml in sampling buffer and 5ng/ml in supernatant of dog faeces suspanded in sampling buffer respectively. Application of this method in detection of coproantigen in 11 experimentally infected dogs proved by arecoline purgation demonstrated 100% positive.