摘要
目的建立一种定量检测乙肝表面抗原(HBsAg)的化学发光免疫分析方法并评估其性能。方法利用双抗体夹心法实验原理,用吖啶酯标记多抗,用生物素标记两株单抗,通过磁颗粒链霉亲和素-生物素系统分离固相和液相完成检测。结果本方法的检测限为0.05 IU/mL,线性范围为0.05~150 IU/mL,可溯源至国际标准品,批内变异小于5%,总变异小于8%,浓度高达106IU/mL的强阳样本未出现HOOK效应;5套阳转盘对雅培Architect、西门子centaur和强生VITROS发光试剂相对敏感系数分别是3、0、-3;国家参考盘检出达标;与雅培Architect比较,2000例临床样本检测灵敏度为99.77%,特异性为100%,一致性达99.95%。结论本研究建立的HBsAg定量检测试剂,各项指标均满足临床检测的要求,灵敏度高,特异性好,适合临床推广应用。
Objective To establish a quantitative detection method for Hepatitis B virus surface antigen(HBsAg)and evaluate its performance.Methods According to the principle of double antibody sandwich,anti-HBsAg polyclonal antibody was linked with acridinium ester,and the other two strains of anti-HBsAg monoclonal antibody were linked with biotin,and magnetic beads were coated by streptavidin.The detection was carried out via streptavidin-biotin separation system and washing process.Results The limit of the detection was 0.05 IU/mL,and the linear range was 0.05-150 IU/mL,also the intra-batch and total variation was lower than 5% and 8%,respectively.No HOOK effect was observed in strong positive samples with a concentration of 10~6 IU/mL.The relative sensitivity coefficients of the 5 seroconvertion panel,compared with Abbott Architect,Siemens Centaur and Johnson VITROS chemiluminescent reagents,were 3,0 and-3,respectively.National reference plate detection results conform to the requirements.Compared with Abbott Architect HBsAg kit,the sensitivity and specificity of the 2000 clinical samples were 99.77%,100%,respectively,and the consistency was 99.95%.Conclusion The HBsAg quantitative test reagent established in this study meets the requirements of the clinical application,with high sensitivity and specificity,and is suitable for clinical application.
引文
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