高灵敏电化学发光法检测人血清中FT4方法的建立  

Establishment of a highly sensitive Electrochemiluminescence Method for the determination of FT4 in human serum

作　　者：席杰 梁辰[1] 邓子辉[1] 冯杰[1] 薛辉[1] 师丹丹 高艳红[1] 颜光涛[1] XI Jie;LIANG Chen;DENG Zi-hui(Department of Clinical Laboratory Center,The First Medical Center of the PLA General Hospital,Beijing 100853,China)

机构地区：[1]解放军总医院第一医学中心医学检验中心,北京100853 [2]深圳普门科技股份有限公司,广东深圳518055

出　　处：《中国实验诊断学》2021年第12期1781-1785,共5页Chinese Journal of Laboratory Diagnosis

基　　金：国家重点研发计划课题子课题(2017YFF0205401)。

摘　　要：目的建立测定人血清游离甲状腺素(FT4)的电化学发光免疫分析方法。方法为检测人血清游离甲状腺素(FT4),将样本和生物素标记的FT4抗原竞争性结合三联吡啶钌标记的抗体,形成的免疫复合物和链霉亲和素包被的磁珠反应,形成了抗原抗体复合物,该免疫复合物通过磁珠吸附到电极上,未结合的物质被清洗掉,随即在三丙胺激发缓冲液的环境下,检测相对光子强度(relative light units,RLU)。根据标准品的RLU数值绘制标准曲线,从而检测出待测样品中的FT4浓度。结果本方法中,最低检测限为0.30pmol/L,批内变异小于10%,批间变异小于15%,准确度为90%-110%,线性范围为0.30-100pmol/L。与罗氏公司FT4检测试剂盒(电化学发光法)平行检测血清样本中FT4浓度,其相关系数为r=0.99(n=44)。将试剂盒经过37℃、8天热处理后,各项指标都达到了标准。结论该方法检测FT4浓度具有高灵敏度、高准确度、线性范围广、稳定性好等优点,所有指标皆符合临床检测的要求。Objective An electrochemiluminescence immunoassay for the determination of free thyroxine(FT4)in human serum was established.Methods The sample and the biotin-labeled FT4 antigen compete for binding to the terpyridine-labeled antibody,the immune Complex combined with streptavidin-coated magnetic particles which formed streptomycin affinity magnetic particle-biotin antigen-ruthenium antibody complex,The composite is adsorbed onto the electrode by magnetic particles,and the unbound material is washed away.Then,tripropylamine challenge buffer was added to detect relative light units(RLU),and a standard curve was established by RLU of the standard to quantitatively measure the content of the sample FT4.Results The method established in this study has a minimum detection limit of 0.30 pmol/L,within batch variation less than 10%,between batch variation less than 15%,and an accuracy of90%-110%,a linear range of 0.30-100 pmol/L.The concentration of FT4 in serum samples was determined in parallel with Roche FT4 detection kit(electrochemiluminescence)with a correlation coefficient of r=0.99(n=44).After the heat treatment at 37℃for 8 days,the performance indicators are up to standard.Conclusion This method has the advantages of wide linear range,high accuracy and good stability,and all the indicators meet the requirements of clinical testing.

关 键 词：FT4 生物素 链霉亲和素 三联吡啶钌 电化学发光法 

分 类 号：R446[医药卫生—诊断学]