高灵敏电化学发光法检测人血清抗甲状腺过氧化物酶抗体

The Establishment of High Sensitive Electrochemical Luminescence Detection Method for Human Serum Anti-TPO

目的通过竞争电化学发光免疫分析的原理,建成一种检测抗甲状腺过氧化物酶抗体(anti-thyroid peroxidase antibody,Anti-TPO)的电化学发光分析方法。方法钌(Ru)标记的Anti-TPO竞争的去结合生物素化的甲状腺过氧化物酶(thyroid peroxidase antibody,TPO),进而和链霉亲和素(streptavidin,SA)包被的磁珠结合,将未结合的钌标记的复合物洗去,含钌并结合SA的复合物吸附到电极上,通过施加电压发光,随后测定RLU(relative light units)。结果本研究建立的Anti-TPO的分析灵敏度(lower limit of detection,LLD,95%置信区间)为5 IU/mL,线性范围为10~600 IU/mL,正确度为0.900~1.100,批间CV<10.0%,批内CV<5.0%。与Roche公司生产的Anti-TPO线性相关系数r=0.9868(n=198),热处理后,试剂性能指标符合要求。结论该方法各项性能指标均能达到临床检测要求,可以推广应用。

Objective Human serum anti-thyroid peroxidase antibody(Anti-TPO)was detected by electrochemiluminescence immunoassay.Methods The specimen,the biotinylated antibody and the terpyridine pyridinium labeled antibody were reacted to form an antigen-antibody sandwich complex,which was combined with streptavidin-coated magnetic particles.The formed complex was washed by magnetic field to remove the unbonded compound.Then tripropylamine excitation buffer was added to detect the relative photon intensity.Results The method established in this study had a minimum detection limit of 5 IU/mL,a linear range of 10-600 IU/mL,and an accuracy of 0.900-1.100,with within batch variation less than 5.0%,between batch variation less than 10%.Compared with Roche company’s Anti-TPO Assay Kit,the correlation coefficient was 0.9868(n=198).After the heat treatment,the performance indicators were up to standard.Conclusion The current method’s various indexes can meet the requirements of clinical testing and can be popularized for future applications.