雌二醇化学发光免疫测定法的建立

Detection of human serum estradiol by chemiluminescent immunoassay

目的 建立灵敏度高 ,测定范围宽的检测人血清雌二醇 (E2 )的化学发光免疫分析法 (CLIA)。方法 采用竞争抑制法 ,用碱性磷酸酶标记抗原 ,金刚烷 (CSPD)增敏化学发光体系作为酶底物。结果 敏感度为 2 .0pg/ml;在 10～ 10 0 0pg/ml之间可以准确定量 ;用不同浓度的E2 质控血清测定精密性 ,批内变异 <8% (n =2 0 ) ,批间变异 <10 % (n =2 0 )。与雌醇、雌酮、雌三醇(E3 )的交叉反应 <1% ,与睾酮、可的松等无交叉反应。试剂具有良好的稳定性 ,在 4℃保存 14个月其整体变化幅度 <10 %。与BeckmanAccessTm及其配套试剂比较有较好的相关性。结论 方法灵敏度高 ,特异性强 ,稳定性好 ,检测范围宽 ,准确性和重复性好。

Objective The purpose of this study is to establish a sensitive chemiluminescent immunoassay (CLIA) with wide linear range for detecting estradiol (E2) in human serum.Methods A competitive binding immunoassay by using direct,chemiluminometric technologe in which an E2-BSA polyclonal antibodies were coated to polystyrene plates.The serum E2 competed with E2-ALP conjugate for the binding sites of anti-E2 antibodies and CSPD was used as.chemiluminescent substrate.Results The lower detection limits of the assay was 10 pg/ml and the upper limit was up to 1000 pg/ml.The assay also offered other advantages,such as easy to use,good precision (intra-assay CV<8% and inter-assay CV<10%=and accuracy.Conclusion Compared with Beckman AccessTM Estradiol,a good correlation between the 2 assays was found.