Bélanger Jean-Christophe, Bandeira Ferreira Fabio Luiz, Welman Mélanie, Boulahya Rahma, Tanguay Jean-François, So Derek Y F, Lordkipanidzé Marie
Montreal Heart Institute Research Center, Montréal, QC H1T 1C8, Canada.
Faculty of Pharmacy, Université de Montréal, Montréal, QC H3C 3J7, Canada.
J Clin Med. 2020 Jan 24;9(2):332. doi: 10.3390/jcm9020332.
The vasodilator-associated stimulated phosphoprotein (VASP) phosphorylation level is a highly specific method to assess P2Y12 receptor inhibition. Traditionally, VASP phosphorylation is analyzed by flow cytometry, which is laborious and restricted to specialized laboratories. Recently, a simple ELISA kit has been commercialized. The primary objective of this study was to compare the performance of VASP assessment by ELISA and flow cytometry in relation to functional platelet aggregation testing by Multiplate whole-blood aggregometry. Blood from 24 healthy volunteers was incubated with increasing concentration of a P2Y12 receptor inhibitor (AR-C 66096). Platelet function testing was carried out simultaneously by Multiplate aggregometry and by VASP assessment through ELISA and flow cytometry. As expected, increasing concentrations of the P2Y12 receptor inhibitor induced a proportional inhibition of platelet aggregation and P2Y12 receptor activation across the modalities. Platelet reactivity index values of both ELISA- and flow cytometry-based VASP assessment methods correlated strongly ( = 0.87, < 0.0001) and showed minimal bias (1.05%). Correlation with Multiplate was slightly higher for the flow cytometry-based VASP assay ( = 0.79, < 0.0001) than for the ELISA-based assay ( = 0.69, < 0.0001). Intraclass correlation (ICC) was moderate for all the assays tested (ICC between 0.62 and 0.84). However, categorization into low, optimal, or high platelet reactivity based on these assays was strongly concordant (κ between 0.86 and 0.92). In conclusion, the consensus-recommended assays with their standardized cut-offs should not be used interchangeably in multi-center clinical studies but, rather, they should be standardized throughout sites.
血管扩张剂相关刺激磷蛋白（VASP）磷酸化水平是评估P2Y12受体抑制作用的一种高度特异性方法。传统上，VASP磷酸化通过流式细胞术进行分析，这种方法费力且仅限于专业实验室。最近，一种简单的ELISA试剂盒已商业化。本研究的主要目的是比较ELISA和流式细胞术评估VASP与通过Multiplate全血凝集法进行的功能性血小板聚集测试之间的性能。将24名健康志愿者的血液与浓度不断增加的P2Y12受体抑制剂（AR-C 66096）孵育。通过Multiplate凝集法以及通过ELISA和流式细胞术进行VASP评估同时进行血小板功能测试。正如预期的那样，P2Y12受体抑制剂浓度的增加在所有检测方式中均引起血小板聚集和P2Y12受体激活的成比例抑制。基于ELISA和流式细胞术的VASP评估方法的血小板反应性指数值高度相关（r = 0.87，P < 0.0001），且偏差极小（1.05%）。基于流式细胞术的VASP检测与Multiplate的相关性（r = 0.79，P < 0.0001）略高于基于ELISA的检测（r = 0.69，P < 0.0001）。所有测试的检测方法的组内相关性（ICC）为中等（ICC在0.62至0.84之间）。然而，基于这些检测方法将血小板反应性分为低、最佳或高的分类具有高度一致性（κ在0.86至0.9 . 2之间）。总之，共识推荐的检测方法及其标准化的临界值在多中心临床研究中不应互换使用，而应在各个研究地点进行标准化。
