Biosystems Data Analysis, Swammerdam Institute for Life Sciences, Universiteit van Amsterdam, Amsterdam, The Netherlands.
Department of Food Science, Chemometrics and Analytical Technology, Faculty of Science, University of Copenhagen, Frederiksberg C, Denmark.
Clin Chem Lab Med. 2019 Dec 18;58(1):103-115. doi: 10.1515/cclm-2019-0729.
Background Characterization of lipoprotein particle profiles (LPPs) (including main classes and subclasses) by means of ultracentrifugation (UC) is highly requested given its clinical potential. However, rapid methods are required to replace the very labor-intensive UC method and one solution is to calibrate rapid nuclear magnetic resonance (NMR)-based prediction models, but the reliability of the UC-response method required for the NMR calibration has been largely overlooked. Methods This study provides a comprehensive repeatability and reproducibility study of various UC-based lipid measurements (cholesterol, triglycerides [TGs], free cholesterol, phospholipids, apolipoprotein [apo]A1 and apoB) in different main classes and subclasses of 25 duplicated fresh plasma samples and of 42 quality control (QC) frozen pooled plasma samples of healthy individuals. Results Cholesterol, apoA1 and apoB measurements were very repeatable in all classes (intraclass correlation coefficient [ICC]: 92.93%-99.54%). Free cholesterol and phospholipid concentrations in main classes and subclasses and TG concentrations in high-density lipoproteins (HDL), HDL subclasses and low-density lipoproteins (LDL) subclasses, showed worse repeatability (ICC: 19.21%-99.08%) attributable to low concentrations, variability introduced during UC and assay limitations. On frozen QC samples, the reproducibility of cholesterol, apoA1 and apoB concentrations was found to be better than for the free cholesterol, phospholipids and TGs concentrations. Conclusions This study shows that for LPPs measurements near or below the limit of detection (LOD) in some of the subclasses, as well as the use of frozen samples, results in worsened repeatability and reproducibility. Furthermore, we show that the analytical assay coupled to UC for free cholesterol and phospholipids have different repeatability and reproducibility. All of this needs to be taken into account when calibrating future NMR-based models.
通过超速离心（UC）对脂蛋白颗粒谱（LPP）（包括主要类别和亚类）进行特征描述具有重要的临床意义。然而，需要快速的方法来替代非常耗时的 UC 方法，一种解决方案是校准快速基于核磁共振（NMR）的预测模型，但 NMR 校准所需的 UC 响应方法的可靠性在很大程度上被忽视了。方法：本研究对 25 个新鲜血浆重复样本的不同主要类别和亚类中的各种基于 UC 的脂质测量值（胆固醇、甘油三酯[TGs]、游离胆固醇、磷脂、载脂蛋白[Apo]A1 和 ApoB）和 42 个健康个体的质控（QC）冷冻混合血浆样本的重复性和再现性进行了全面研究。结果：胆固醇、ApoA1 和 ApoB 的测量在所有类别中都具有非常高的重复性（组内相关系数[ICC]：92.93%-99.54%）。主类和亚类中的游离胆固醇和磷脂浓度以及高密度脂蛋白（HDL）、HDL 亚类和低密度脂蛋白（LDL）亚类中的 TG 浓度的重复性较差（ICC：19.21%-99.08%），这归因于低浓度、UC 过程中引入的变异性和测定方法的局限性。在冷冻 QC 样本中，发现胆固醇、ApoA1 和 ApoB 浓度的重现性优于游离胆固醇、磷脂和 TG 浓度。结论：本研究表明，对于某些亚类中接近或低于检测限（LOD）的 LPPs 测量值，以及使用冷冻样本，会导致重复性和再现性变差。此外，我们还表明，与 UC 结合使用的游离胆固醇和磷脂分析测定具有不同的重复性和再现性。在对未来基于 NMR 的模型进行校准时，需要考虑到所有这些因素。
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