ELISA检测人IgG确定种属的改良方法——酶联免疫在物证检验中的应用研究之二

本实验采用含1/10,000戊二醛的0.05M,pH9.6碳酸盐缓冲液稀释抗体包被反应板,并使酶标记抗体浓度由1/100提高到1/50,使实验时间由常规方法的20多小时缩短到6小时。1∶320,000稀释的标定的人新鲜血痕浸液仍能得出阳性结果。本实验除用聚苯乙烯反应板外,还使用了聚氯乙烯反应板。两种反应板的实验条件及结果相同。聚氯乙烯反应板可以根据实验所需反应孔的多少进行剪裁,从而使实验材料得到节约。这两种反应板都可以预先用抗体包被后,置-20℃冰箱保存备用,从而简化了操作。本方法快速、灵敏、简单、重复性好,便于推广。

A MODIFIED METHOD OF DETECTION OF HUMAN IgG FOR DETERMINATION OF BLOODSTAINS SPECIES USING ELISA

A modified method reported in this paper is based on our last report identification of species of bloodstains by detecting human IgG using ELISA. In this method the coating buffer contained 1/10,000 glutaraldehyde was used, and the concentration of the labelled-antibody was raised correspondingly. The microtiter wells were coated with antibody for 2 hours, and the antigen reacted for 1 hour, the labelled-antibody for 1 hour, the substrate for 1/2 hour. The test time could be shortened from routine 20 hours to 6. The sensitivity of this method was over 1∶320,000 for the dilution of the standarded human fresh bloodstains extracts and other indexes were as satisfied as last report. Both of the polystyrene and the polyvinyl chloride microtiter plates were used under the same condition, and got the same results. The latter plate could be cut on the basis of needs. Two kinds of plates could be coated with anti-HIgG in advance and stored at -20℃ for use.