目的 使用抗-异硫氰酸荧光素(FITC)固相包被板建立促黄体生成素(LH)的酶联免疫(ELISA)检测方法.方法用FITC及辣根过氧化物酶(HRP)分别标记两株抗-LH单克隆抗体,建立一步检测LH的ELISA检测方法,并与经典的双抗体夹心法进行了方法学对比评价.结果 FITC与抗-FITC系统检测LH,在2~50mIU/mL的校准曲线范围内相关系数0.9968,分析内精密度为7.6%,分析间精密度为7.02%,热稳定性下降18.5%,与双抗体夹心法相当,空白检测限为0.15mIU/mL,优于双抗体夹心法,钩状效应(HOOK效应)比双抗体夹心法差.与贝克曼检测结果回归方程Y=0.970X+0.614,相关系数r=0.975.结论 成功建立基于FITC与抗-FITC系统的ELISA检测方法,与双抗体夹心法相比,两种方法均能满足临床检测的需要.%Objective To develop an enzyme-linked immunosorbent assay (ELISA) for the detection of luteinizing hormone (LH) using an anti-FITC solid-phase coating plate.Methods Two anti-LH monoclonal antibodies were labeled with fluorescein isothiocyanate (FITC) and horseradish peroxidase (HRP), respectively.The ELISA method was established to detect luteinizing hormone (LH) in one step.The classic method of double antibody sandwich method was compared and evaluated.Results The correlation coefficient of FITC and anti-FITC system was 0.9968 within the range of 2-50mIU/mL.The precision of the assay was 7.6% and the inter-analytical precision was 7.02%.The thermal stability was decreased by 18.5%.The blank detection limit was 0.15mIU/mL, which was better than double antibody sandwich method.Meanwhile, the HOOK effect was worse than double antibody sandwich method.The correlation equation was Y=0.970X+0.614, and the correlation coefficient was r=0.975.Conclusion ELISA method based on FITC and anti-FITC system was successfully established.Compared with double antibody sandwich assay, we found both methods can meet the need of clinical detection.
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