酶标法检测TSGF不确定度的评估分析

The evaluation of measurement uncertainty of TSGF with ELISA

目的:探讨实验过程中不同因素对定量酶联免疫吸附试验(EL ISA)检测结果的影响,评估ELISA方法检测TS-GF的不确定度。方法:参考CNAS-GL06:2006《化学分析中不确定度的评估指南》等测量不确定度指南文件,重复检测低、中、高值三个混合标本,对本实验室ELISA法检测肿瘤相关物质TSGF的测量不确定度进行评估。结果:低、中、高值标本测定结果分别为52.90 U/ml、60.04 U/ml、78.75 U/ml,其相应的扩展不确定度为10.32 U/ml、11.63 U/ml、15.16 U/ml。移液器加样产生的不确定度所占比例最高,酶标仪测定OD值引入的不确定度所占比例最小。随着标本浓度的上升,不确定度逐渐增大,精密度在总不确定度中所占比例逐渐升高,与此相反,标准曲线拟合引入的不确定度随标本浓度升高逐渐降低。结论:在临床工作中应严格控制ELISA实验条件,特别是控制室温和加样过程,尽量保证实验操作的准确性和标准曲线的稳定性,以获得准确可靠的结果。

Objective：To investigate the effects of different factors on the results of quantitative ELISA,and evaluate the measurement uncertainty of ELISA taking Tumor Supplied Group of Factors（TSGF） as an example.Methods： Referred to CNAS-GL06： 2006 ＂Guidance on Evaluating the Uncertainty in Chemical Analysis＂ and other guidelines,three samples of low value,middle value and high value were repeatedly measured,and the measurement uncertainty of ELISA was estimated.Results： The results of the three samples were 52.90 U/ml,60.04 U/ml,78.75 U/ml,and the expanded uncertainty were 10.32 U/ml,11.63 U/ml,15.16 U/ml.The uncertainty ratio of adding sample of transferpettor was the highest,and the uncertainty ratio of the value of absorbance was smallest.With the sample value increasing the uncertainty of the precision increased gradually,but the uncertainty of the calibrator curve decreased.Conclusion： To gain accurate and reliable results,the conditions of ELISA tests,in particular,the room temperature and adding sample process should be controlled strictly and make sure of the stability and accuracy of operation and the standard curve.