硫酸去氢表雄酮定量测定试剂盒(化学发光免疫分析法)的研制及性能评价

Development and evaluation of a quantitative assay for dehydroepiandrosterone sulfate: chemiluminescence immunoassay

目的研制硫酸去氢表雄酮定量测定试剂盒(化学发光免疫分析法),并对其试剂盒性能进行评价。方法利用竞争抑制法,对原料进行筛选,建立硫酸去氢表雄酮定量测定试剂盒。并分别对本试剂盒的准确性、最低检测限、线性、重复性、特异性、参考范围及样本对比方面进行评价。结果经过筛选,选用反应模式:磁微粒包被羊抗鼠Ig G-鼠抗硫酸去氢表雄酮单克隆抗体-吖啶酯标记DHEAS-BSA;对此方法建立的试剂盒进行方法学评价,最低检测限可达1.84μg/d L,线性在0~1 500μg/d L之间,相关系数r=0.999 8,准确度的回收率为105.38%,重复性均小于8%;建立了健康人参考范围(女性:47.9~407.5μg/d L,男性97.1~522.5μg/d L),与西门子ADVIA Centaur测定结果显著相关,线性方程为y=1.078 13+0.992 16x,相关系数r=0.994 7,2种试剂盒的测定结果具有较高的一致性。结论本研究建立的硫酸去氢表雄酮定量测定试剂盒(化学发光免疫分析法)各项性能均达到了临床检验的要求,为国产化66硫酸去氢表雄酮定量测定试剂盒(化学发光免疫分析法)的研发奠定了基础,有望用于临床血清硫酸去氢表雄酮的检测。

Objective To develop and evaluate the performance of the Dehydroepiandrosterone sulfate quantitative determination kit： chemiluminescent immunoassay. Methods The method of competitive inhibition was used to screen the raw materials and to establish the Dehydroepiandrosterone sulfate quantitative determination kit with chemiluminescent immunoassay. The performance of accuracy, limit of detection, linearity, repeatability, analytical specificity, reference interval and samples comparison were determined. Results After screening, the reaction mode of magnetic particle coated goat anti mouse Ig G-mouse anti DHEAS antibodyacridine ester labeled DHEAS-BSA was selected. The limit of detection was less to 1.84 μg/d L. This assay was designed to be linear across the measurement range of 0-1 500 μg/d L with value r=0.999 8. The recovery rate was 105.38%. The repeatability was less than 8%. The DHEAS reference interval of healthy population was established（female： 47.9-407.5 μg/d L, male： 97.1-522.5 μg/d L）. The determination results of serum samples were significantly correlated with results tested by SIEMENS ADVIA Centaur assay with the linear equation was y=1.078 3 ＋ 0.992 16 x and the value r=0.994 7. The results of these 2 assays showed high consistency.Conclusion All the performance characteristics of this kit reached the requirements for clinical detection and laid a foundation for establishing the domestic quantitative assay of dehydroepiandrosterone sulfate, which is able to apply to the clinical detemination of serum DHEA-S.