血清铁蛋白化学发光免疫分析方法的建立及临床应用

Construction and Clinical Application of Chemiluminescent Immunoassay for Human Serum Ferritin

摘要:

利用链霉亲和素-生物素放大系统及双抗夹心法，建立检测血清铁蛋白的化学发光免疫分析方法。将生物素、辣根过氧化物酶分别标记铁蛋白的一对抗体，以链霉亲和素包被96孔发光板，铁蛋白标准品与国际标准品进行溯源，并对本法的各项性能指标进行评价。收集300份临床血清样本，使用本法与进口试剂盒同时检测，检测结果显示：相关系数r为0.97，本法的线性测量范围为10~800 ng/mL，灵敏度为0.2 ng/mL，批内、批间的变异系数（CV%）分别为3.4%~5.5%及5.0%~7.3%，与癌胚抗原、人白蛋白、人类心脏铁蛋白以及血红蛋白交叉反应率均小于1%。本法37 ℃、9 d热稳定性良好。表明所建立的方法各项指标均满足临床检测要求，抗体及样本用量少，操作简便，反应迅速，便于临床应用，可替代国外同类试剂盒。

Abstract:

To develop a chemiluminescent immunoassay method for detecting serum ferritin with biotin-avidin amplified system and a sandwich way. A pair of antibodies were labeled with biotin and horseradish peroxidase respectively. The 96 well microplate was coated with streptavidin. The serum ferritin standard and international standard was evaluated. 300 clinical serum specimens wer detected with this method and a foreign serum ferritin kit（Roche），the correlation coefficient was 0.97. The linear measurement range of this method was 10~800 ng/mL. The intra-assay and inter-assay coefficients of variation were 3.4~5.5%， and 5.0~7.3%. The cross-reactivity rate with CEA（carcinoembryonic antigen），human albumin，human heart ferritin，and hemoglobin were all less than 1%. The developed method was stable 9 days at 37 ℃. This assay was easily operable，small amount of specimens needed and the reaction was quick，so it was suitable for clinical application.