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.
作者
席杰
梁辰
邓子辉
冯杰
薛辉
师丹丹
高艳红
颜光涛
XI Jie;LIANG Chen;DENG Zi-hui(Department of Clinical Laboratory Center,The First Medical Center of the PLA General Hospital,Beijing 100853,China)
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)。
作者简介
通讯作者:颜光涛。
