目的:血清淀粉样蛋白A(SAA)和乳铁蛋白(LTF)的临床价值已受到广泛关注。但是他们的检测方法不足,这限制了它的应用。本研究旨在开发一种基于稳定元素标记策略和电感耦合等离子体质谱(ICP-MS)的SAA/LTF双重检测方法,并评估其是否可以在临床实践中广泛使用。
方法:构建了基于夹心法的双重免疫测定系统。优化后,按照临床实验室标准研究所(CLSI)的指南进行方法学评价.最后,收集131份血浆样本,分析新方法是否适用于临床检测。
结果:LoB,LLoQ,ULoQ,测定的线性范围为1.09ng/mL,3ng/mL,1500ng/mL,3-1500ng/mL的SAA和0.85ng/mL,2ng/mL,1200ng/mL,对于LTF分别为2-1200ng/mL。回收率为95.01%~106.26%,批次内精度低,中间,高水平样本<8%,它们的批次间<11%,干扰试验结果偏差小于±10%。SAA的曲线下面积(AUC)为0.9809,LTF为0.8599,和0.9986的组合。
结论:SAA/LTF的双重定量免疫测定具有较高的准确性,精度好,和高特异性,符合临床检测要求,可广泛应用于临床。
OBJECTIVE: The clinical value of Serum amyloid A (SAA) and Lactoferrin (LTF) has received significant attention, but their detection methods are inadequate, which limits their application. This study aims to develop a dual detection method based on stable element labeling strategies and inductively coupled plasma mass spectrometry (ICP-MS) for SAA/LTF and to assess whether it can be widely used in clinical practice.
METHODS: A duplex immunoassay system based on sandwich method was constructed. After optimization, methodological evaluation was performed with the guidelines of Clinical Laboratory Standards Institute (CLSI). Finally, 131 plasma samples were collected to analyze whether the new method is suitable for clinical detection.
RESULTS: The LoB, LLoQ, ULoQ, and linear range of the assay were 1.09 ng/mL, 3 ng/mL, 1500 ng/mL, 3-1500 ng/mL for SAA and 0.85 ng/mL, 2 ng/mL, 1200 ng/mL, 2-1200 ng/mL for LTF respectively. The recovery rates were 95.01% to 106.26%, the intra-batch precision of low, intermediate, and high-level samples was <8%, and the inter-batch of them was <11%, the deviation of interference test results was less than±10%. The Area Under the Curve (AUC) was 0.9809 for SAA, 0.8599 for LTF, and 0.9986 for combination.
CONCLUSIONS: The quantitative duplex immunoassay for SAA/LTF has high accuracy, good precision, and high specificity, which meets the clinical testing requirements and can be widely used in clinical practice.