高灵敏化学发光法特异性检测人血清促甲状腺激素方法的建立  

The Development of a Highly Sensitive Electrochemiluminescence Method for the Accurate Quantification of Thyroid Stimulating Hormone in Human Serum

作　　者：冯杰[1] 高艳红[1] 井三有 赵强 杨锐创 颜光涛[1] FENG Jie;GAO Yanhong;JING Sanyou;ZHAO Qiang;YANG Ruichuang;YAN Guangtao(Department of Laboratory Medicine,the First Medical Center,Chinese PLA General Hospital,Beijing 100853,China;Shenzhen Mindray Bio-Medical Electronics Co.,Ltd,Shenzhen 518063,China)

机构地区：[1]解放军总医院第一医学中心检验科,北京100853 [2]深圳迈瑞生物医疗电子股份有限公司,广东深圳518063

出　　处：《标记免疫分析与临床》2024年第1期157-161,共5页Labeled Immunoassays and Clinical Medicine

基　　金：国家重点研发计划项目子课题(编号:2017YFF0205401)。

摘　　要：目的 通过双位点夹心化学发光免疫分析原理,建立一种高灵敏度测定人血清促甲状腺激素(thyroid stimulating hormone, TSH)的化学发光免疫分析方法。方法 将样本与预包被有抗促甲状腺激素(TSH)抗体的超顺磁珠,以及抗TSH-碱性磷酸结合物添加到反应管中共孵育,待测样本中的TSH通过夹心法与磁珠和抗TSH-碱性磷酸结合物与形成双位点复合物。与磁珠结合的复合物吸附到磁场上,用缓冲液洗去未结合的物质。加入发光底物3-(2-螺旋金刚烷)-4-甲氧基-4-(3-磷氧酰)-苯基-1,2-二氧环乙烷二钠盐[3-(2-spiroadamantane)-4-methoxy-4-(3-phosphoryloxy)-phenyl-1,2-dioxetane, AMPPD],测定相对光子强度(RLU),通过标准曲线计算待测样本中TSH的浓度。结果 本研究建立的TSH检测方法,最低检测限为0.003μIU/mL,批内CV小于1%,总CV小于3%,正确度样本实测值与靶值之间相对偏差均在±12.5%范围内,线性范围为0.005~100μIU/mL且相关系数(r)高于0.9900。开封稳定性试验的相对偏差<10%。结论 该方法检测血清TSH浓度具有灵敏度高、准确性好、线性范围广、稳定性强的优点,各项指标均达到临床检测要求,可以推广使用。Objective To develop a high-sensitivity chemiluminescence immunoassay method for the determination of thyroid stimulating hormone(TSH)in human serum based on the principle of double-site sandwich chemiluminescence immunoassay.Methods The study samples were co-incubated with superparamagnetic beads pre-coated with anti-thyrotropin antibody(TSH)and anti-TSH-alkaline phosphate conjugate in the reaction tube.TSH in the sample was combined with magnetic beads and anti-TSH-alkaline phosphate conjugate by sandwich method to form a two-site complex.The complex bound to the magnetic bead was adsorbed to the magnetic field and the unbound material was washed away with a washing buffer.The luminescent substrate,3-(2-spiroadamantane)-4-methoxy-4-(3-phosphoryloxy)-phenyl-1,2-dioxetane(AMPPD)was added,and the relative photon intensity(RLU)was measured.The concentration of TSH in the sample was calculated using a standard curve.Results TSH detection method established in this study showed that the minimum detection limit was 0.003μIU/mL,in-batch CV was less than 1%,and total CV was less than 3%.The relative deviation between the measured value and the target value of the sample was within the range of±12.5%,while the linear range was from 0.005 to 100μIU/mL,and the correlation coefficient(r)was higher than 0.9900.The relative deviation of the stability was less than 10%.Conclusion The proposed method in this study demonstrates a high sensitivity,high accuracy,wide linear range and good stability for quantifying serum TSH,and all indicators meet the requirements of clinical testing and can be popularized for future applications.

关 键 词：促甲状腺激素 化学发光免疫分析法 灵敏度 

分 类 号：R-331[医药卫生]