细粒棘球蚴囊液抗原纯化方法的比较评价

对于绵羊肝脏细粒棘球蚴囊液用常规粗制法,通过抗绵羊全血清抗体免疫吸附柱的亲和层析法,Oriol氏纯化法,Burstein氏纯化法,以及将纯化抗原再经酚提取法制备的六种抗原,以SDS-PAGE,免疫电泳,ELISA,IHA等方法进行了抗原得率、纯度、组份、免疫活性和特异性等方面的比较。结果表明:亲和层析抗原得率最高(l6.25％～22.64％),Oriol氏法纯化抗原最低(3.35％)。与囊液粗抗原相比,纯化抗原中Burstein抗原组份最多。经酚提取的纯化抗原组份最少。各种抗原组份数目显然相差明显,但在免疫电泳中均出现弧5沉淀线。酚提取法不能达到将抗原5和抗原B分离的目的。在ELISA和IHA中,Burstein法纯化抗原的活性最高。作者分析了亲和层析抗原活性低于Burstein和Oriol两种抗原的原因可能是亲和层析抗原缺少68kDa和36.5kDa两种宿主组份。因而认为这两个组份可能同时具有寄生虫抗原的性质。在ELISA试验中,各种纯化抗原均与囊虫病人血清产生部分交叉反应,但比粗抗原低。在IHA试验中,纯化抗原均不与囊虫病人血清产生交叉反应。作者推荐由于Burstein法纯化抗原得率高,提纯程序较简单,抗原活性高而且在IHA中没有与囊虫病人血清的交叉反应,因而可作为常规诊断抗原使用。

A Comparative Evaluation of Antigens from Sheep Hydatid Cyst Fluid Purified by Different Methods

A comparative study on the antigens of Echinococcus granulosus prepared from sheep liver hydatid cyst fluid with different methods, including affinity chromatography, Oriot's mehod, Burstein's method and phenol abstraction, was carried out. The antigens were compared from aspects of yield, purity, fraction,immunoactivity and specificity. The results suggested that antigen yield was highest (16.25%-22.64%) by affinity chromatography method and lowest by Oriol's method (3.35%). The fractions (bands in SDS-PAGE) were maximum in the antigen purified by Burstein's method and minimum by phenol extraction. Although fractions were obviously different, the arc 5 occured in all of the antigens by immunoelectrophoresis. It was revealed that the phenol fraction method could not isolate antigen 5 and antigen B. The immunoactivity of Bursten's antigen was highest in ELISA and IHA. The reason why the immunoactivity of antigen purified with affinity chromatography is lower than Burstein's and Oriol's antigens is discussed. It is probably for the lack of 68kD and 36.5kD fractions of the host in affinity chromatography antigen while the other two fractions may have both the host and parasite nature. All of the antigens were reacted with sera from cysticercosis patients in ELISA but showed no cross reaction in IHA. The authors recommand that the Burstein's antigen may be used in serodiagnosis of hydatid disease as choice antigen before ideal antigens are available.