促甲状腺激素液相芯片检测法的建立和方法学评价

Development of a liquichip assay for thyrotropic-stimulating hormone and evaluation of the methodology

目的建立用液相芯片技术测定促甲状腺激素（TSH）的方法，对其方法学和临床应用价值作评价。方法通过抗TSH的单抗交联单色荧光微球，采用标记有生物素的另一配对抗TSH的单抗以及由藻红蛋白（PE）标记的亲合素组成相应的荧光报告系统，用液相芯片仪Luminex100检测待检标本荧光强度，通过TSH标准浓度绘制出标准TSH浓度荧光强度剂量曲线。用液相芯片技术测定100份临床血清标本的TSH浓度并与拜尔的化学发光法的测定结果做方法学比较。结果液相芯片技术测定TSH的线性范围为0．3～162μIU／mL，高、中、低3个浓度的批内精密度为10．00％、6．91％、17．80％。高、中、低3个浓度的回收率为94．3％、96．7％、95．2％。与拜尔的化学发光法的相关系数为0．965（P=8．77×10^-57）。结论本研究建立的液相芯片技术在TSH测定中的主要技术指标与拜尔的化学发光法相近，有临床应用价值。

Objective To develop liquichip assay for thyrotropic-stimulating hormone （TSH） , and evaluate the method and the value of clinical application. Methods Monoclonal antibodies of TSH were coupled to fluorescent beads produced by Luminex corporation. Fluorescent assay system was formed by streptavidin R-phycoerythrin and the other TSH biotinylated monoclonal antibodies which were pared with the antibodies coupled to beads. Dose-responses curve was plotted with standard solutions of TSH. The Luminexl00 was used to measure fluorescent intensity for analyzing the concentrations of TSH in 100 clinical serum samples and compared with Bayer chemiluminescence immunoassay for TSH assay. Results Analytical measurement range of the method was from 0.3 to 162 μIU/mL. Reproducibility coefficient of variation were 10.00% , 6.91% and 17.80% in high, medium and low TSH concentrations respectively. Recovery rates were 94.3% , 96.7% and 95.2% in high, medium and low TSH concentrations respectively. Coefficient of Pearson was 0. 965 in correlation analysis with Bayer chemiluminescence immunoassay （P = 8.77 ×10^-57 ）. Conclusions The major technical indexes of the liqnichip assay are almost equal to Bayer chemiluminescence immunoassay and it is worthwhile for clinical application.