己烯雌酚酶联免疫检测方法的建立

建立灵敏度高和特异性强的己烯雌酚残留检测分析方法。用人工合成的己烯雌酚免疫原多次免疫家兔,获取高效价抗体。纯化抗体经生物素标记,可同辣根过氧化物酶(HRP)标记的亲和素特异结合。采用纯化的抗体包被96孔板,形成固相抗体。加入不同浓度的己烯雌酚标准品后,再加入生物素标记抗体,以酶标记亲和素作为探针,经酶底物显色反应可获得良好的ELISA标准曲线。该方法检测灵敏度为0.125 ng/mL,可定量检测范围为0.25~15.60 ng/mL。该方法操作简便,具有较强灵敏度和特异性,可满足实践中一般样品的残留检测。

Establishment of ELISA for Diethylstilbestrol

To develop a high sensitive and high specific ELISA for DES.The high effect antibody was obtained by immunizing rabbits with recombinant DES more times.The purified anti-DES IgG was labeled with biotin and then specially linked to avidin which was labeled with HRP.The purified antibody was used to coat 96 hole board as capture antibody.After adding different concentration of DES standards,then biotin-IgG,avidin-HRP were added in turn to the reaction system.DES levels were determined by the color,and standard curve was obtained.The sensitivity of this method was 0.125 ng/mL and the measuring range was 0.25-15.60 ng/mL.This is one of simple,sensitive and specific method for measurement of DES,it can be used to measure DES level in practice.