游离β人绒毛膜促性腺激素定量测定化学发光免疫分析法的建立与性能评价

Quantitative determination of free β human chorionic gonadotropin:establishment and performance evaluation of chemiluminescence immunoassay

目的建立一种定量测定游离β人绒毛膜促性腺激素(freeβhuman chorionic gonadotrophin,f-β-h CG)的化学发光免疫分析法,并对其性能进行评价。方法利用双抗体夹心法,对反应步骤进行筛选,建立游离β人绒毛膜促性腺激素定量测定的化学发光免疫分析法,并对其最低检测限、准确度、重复性、批间精密度、特异性、参考范围及样本比对进行评价。结果经过筛选,选用了二步法反应步骤;对f-β-h CG试剂盒进行方法学评价,最低检测限可达0.006 ng/m L,准确性相对偏差在2.56%～4.43%,线性在0～200 ng/m L,相关系数R^2=1,重复性CV值均小于8%,批间CV值均小于15%;与罗氏的游离β-绒毛膜促性腺激素定量测定试剂盒(电化学发光法)测定结果显著相关,一致性达0.965,线性回归方程为y=1.131x+0.826,相关性r值为0.992,2种试剂盒的检测结果具有较高的一致性。结论本研究自制试剂盒的各项性能均能符合临床检验的要求,有望能替代昂贵的进口试剂应用于临床检测及研究。

Objective To establish a chemiluminescence immunoassay for quantitative determina-tion of freeβhuman chorionic gonadotrophin(f-β-hCG)and evaluate its performance.Methods The dou-ble antibody sandwich method was used to screen the reaction steps,and the chemiluminescence immunoassay method for quantitative determination of freeβhuman chorionic gonadotropin was established.The minimum detection limit,accuracy,repeatability,inter-assay precision and specificity were determined.The reference range and sample comparison were evaluated.Results After screening,the two-step reaction step was selected.The methodological evaluation of f-β-hCG kit was carried out.The minimum detection limit was 0.006 ng/mL,the relative deviation of accuracy was 2.56%-4.43%,the linearity was 0-200 ng/mL,the correla-tion coefficient R2=1,the repetitive CV values are less than 8%,and the inter-assay CV values are less than 15%.Compared to the Roche's freeβ-chorionic gonadotropin quantification kit(electrochemiluminescence method),the results were significantly correlated,the consistency was 0.965,the linear regression equation was y=1.131x+0.826,and the correlation r value was 0.992.The detection results of the 2 kits were highly consistent.Conclusion The performance of the home-made kits in this study can meet the requirements of clinical testing,and it is expected to replace expensive imported reagents for clinical testing and research.