对硫磷ELISA异源分析模式比较研究

为探讨方法模式对异源ELISA法各分析参数的影响情况及影响机理,设计并合成了4种与对硫磷结构相似的竞争半抗原,通过建立不同模式的同源与异源ELISA分析法,比较了不同模式下的抗原抗体用量、灵敏度、检测限及不同样品添加回收率。结果表明,异源分析对抗原和抗体的用量比同源分析有所增加,但异源分析可大大提高方法的检测灵敏度。在异源分析中,直接竞争包被抗原的ELISA(CC模式)方法重现性较差;直接竞争包被抗体的ELISA(AC模式)抗原抗体用量最多,检测灵敏度较低;间接竞争ELISA法(IC模式)具有较好的检测灵敏度,抗环境和基质干扰的能力较强,为异源ELISA分析中较优的模式。

Comparative Study of Three ELISA Formats forHeterologous Assay of Parathion

Four competitors which simulate parathion molecule were designed and synthesized,followed by testing the effect of assay format on the heterologous ELISA.Three types of assay formats,named the conjugate-coated (CC) format,antibody-coated (AC) format and indirect competitive (IC) format,were applied in the homologous or heterologous ELISA.Some factors,such as dosages of haptens and the antibody mentioned above,IC₅₀ value and limit of detection of parathion,and recoveries of ELISA in different samples were compared to find the best assay format.The results showed that the used dosage of hapten and antibody in the heterologous ELISA were higher than that in the homologous ELISA,the heterologous ELISA greatly improved the sensitivity.Moreover,in the heterologous ELISA,the CC format had a relatively low reproducibility.While the AC format had the highest dosage of hapten and antibody in the three types formats,but had low sensitivity.It's found that the IC format had a preferable sensitivity and stable resistance to matrix effects.In conclusion,the IC format was the most suitable one in the heterologous ELISA.