微囊藻毒素的酶联免疫法分析影响因素探讨

考察样品与抗微囊藻毒素(MC)-LR的单克隆抗体(单抗)加入间隔时间、温育温度、检测时间、pH、H2O2、Fe2+等不同因素对酶联免疫法(ELISA)测定MC-LR的影响。结果表明:(1)随着样品与单抗加入间隔时间延长,MC-LR测定值逐渐降低。(2)25、37℃下,MC-LR测定值几乎没有差别,测定结果与取用标准样品具有很好的一致性。(3)不同检测时间下,无论是低浓度还是高浓度标准样品,MC-LR测定值均有一定变化,虽然幅度均不大,但仍然建议终止反应后尽快比色。(4)当pH为7～9时,MC-LR测定值变化不大。(5)当H2O2摩尔浓度为0.1、1.0、10.0mmol/L时,均会对ELISA产生干扰。通过水浴去除H2O2后,H2O2初始摩尔浓度分别为0.1、1.0mmol/L时,MC-LR回收率分别达到87%及81%。(6)Fe2+对酶有显著的抑制作用,可通过沉淀过滤的方式消除干扰。

Study on the influential factors in the detection of microcystins by enzyme linked immunosorbent assay

The enzyme linked immunosorbent assay(ELISA) was employed to detect microcystins(MC).The effect of MC-LR and it’s monclonal antibody addition intervals,incubation temperature,detection time,pH,H2O2 and Fe2+ on the detection results was investigated.Results showed that the MC-LR detection value was decreased with increasing the MC-LR addition intervals.When the incubation temperatures was 25,37 ℃,the MC-LR detection value was almost the same;the determination results and standard sample had very good consistency.The MC-LR determination value had certain change for both low concentration and high concentration standard samples,although the variation was not big,it was still recommend to carry the colorimetric analysis as soon as possible.pH(7-9) had little effect on ELISA;ELISA was disturbed when H2O2 was 0.1,1.0,10.0 mmol/L.The recovery of microcystins increased significantly after H2O2 removal,when the H2O2 was 0.1,1.0 mmol/L,the microcystins recovery rate was 87% and 81% respectively.Fe2+ had significant inhibition on ELISA.