吖啶酯直接标记测定血清总T3化学发光免疫分析方法的建立和初步验证  

An Acridinium Ester Labeled Chemiluminescence Immunoassay for the Quantitative Determination of Total T3 in Human Serum

作　　者：吕东川 贺敏 LYU Dong-chuan;HE Min(Beijing North Institute of Biological Technology Co.,Ltd.,Beijing100076,China)

机构地区：[1]北京北方生物技术研究所有限公司

出　　处：《标记免疫分析与临床》2019年第12期2135-2140,共6页Labeled Immunoassays and Clinical Medicine

摘　　要：目的建立以吖啶酯为标记物检测三碘甲状腺原氨酸(T 3)的磁微粒化学发光免疫分析方法并初步验证该方法作为体外诊断试剂应用的能力。方法通过二抗将抗-T 3抗体固定在磁微粒上,比较了平衡竞争法和顺序加样法两种反应模式,用最终选定的条件对该方法的精密度、定量检出限、回收率、线性范围、抗干扰能力进行了验证,并与进口全自动化学发光试剂检测结果进行了比对。结果本方法最终采用顺序加样法的反应模式,高、低水平两份样本变异系数分别为4.0%和3.9%,0.4nmo/L的样本满足定量检出限的要求,两份样本的回收率分别为99.8%和105.1%,样本水平在0.5~12nmol/L的范围内满足线性要求,血红蛋白<2.5mg/mL、胆红素<0.3mg/mL、三酰甘油<10mg/mL、胆固醇<10mg/mL时对血清样本测量无干扰。与进口全自动化学发光试剂同时测定69份血清样本,二者测定结果的相关系数r为0.992。结论成功建立了血清总T 3化学发光免疫分析法,可以满足临床诊断的要求。Objective To establish a magnetic particle chemiluminescence immunoassay method for the determination of triiodothyronine(T 3)with acridinium ester as a marker and to verify its application as an in vitro diagnostic reagent.Methods Anti-T 3 was immobilized on magnetic particles.The two reaction modes of equilibrium competition and sequential loading were compared for their performances.The repeatability,LoQ and recovery rate of the method were determined by the final selected conditions.The linear range and interference testing results were verified.Results The study adopted the reaction mode of sequential loading method as the final model.The within-laboratory CV were 4.0%and 3.9%,respectively.The sample of 0.4nmo/L met the requirement of LoQ.The recovery rate of the two samples were 99.8.%and 105.1%,respectively,and sample concentration in the range of 0.5-12nmol/L met linear requirements.Within the ranges of Hemoglobin<2.5mg/mL,bilirubin<0.3mg/mL,triglyceride<10mg/mL,and cholesterol<10mg/mL,results showed no significant interference with the measurement.Conclusion The CLIA system for detecting serum total T 3 has been established,which can meet the requirements of clinical diagnosis.

关 键 词：三碘甲状腺原氨酸 吖啶酯 全自动化学发光 

分 类 号：O65[理学—分析化学]