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一种基于酶联免疫吸附测定法的新型血管舒张剂相关刺激磷蛋白磷酸化检测方法在评估中国人群血小板反应性指数中的验证

Validation of a New ELISA-Based Vasodilator-Associated Stimulated Phosphoprotein Phosphorylation Assay to Assess Platelet Reactivity Index in a Chinese Population.

作者信息

Ding Peng, Wei Yujie, Chen Nana, Liu Huiliang

机构信息

1 Chinese People's Liberation Army General Hospital, Beijing, China.

2 Department of Cardiology, General Hospital of Chinese Armed Police Force, Beijing, China.

出版信息

Clin Appl Thromb Hemost. 2018 Apr;24(3):452-461. doi: 10.1177/1076029616689300. Epub 2017 Jan 23.

Abstract

The level of platelet reactivity during P2Y12-adenosine diphosphate receptor antagonist is associated with ischemic and bleeding risks following percutaneous coronary intervention in acute coronary syndrome. Determining platelet reactivity inhibition may be valuable for confirming effective platelet inhibition for individual patients and identifying patients at risk of bleeding. The enzyme-linked immunosorbent assay (ELISA)-based vasodilator-stimulated phosphoprotein (VASP) assay offers unique advantages over other methods and has not been used in the Chinese population. We enrolled 10 healthy volunteers and 54 patients with acute coronary syndrome. The volunteers received no treatment, and patients were administered a loading dose of clopidogrel or ticagrelor. The platelet reactivity index (PRI) was measured using flow cytometry (FC)-VASP and ELISA-VASP at baseline and 8-hour postloading dose. Blood samples of healthy volunteers and clopidogrel- or ticagrelor-treated patients were frozen and stored for 1, 2, and 4 weeks after initial activation. All frozen samples were tested using ELISA-VASP. The PRI assessed by FC-VASP and ELISA-VASP correlated well showing a high degree of consistency in identifying high or low on-treatment platelet reactivity. No significant time-dependent changes in PRI results were observed in frozen samples stored up to 4 weeks compared to nonfrozen samples. The PRI of ticagrelor-treated patients was lower than that of clopidogrel-treated patients. The ELISA-VASP effectively assesses the PRI, and results obtained from frozen specimens are unaffected by storage and shipment prior to assay. Ticagrelor was superior to clopidogrel in decreasing the PRI.

摘要

急性冠状动脉综合征患者经皮冠状动脉介入治疗后,P2Y12-二磷酸腺苷受体拮抗剂治疗期间的血小板反应性水平与缺血和出血风险相关。确定血小板反应性抑制对于确认个体患者的有效血小板抑制以及识别出血风险患者可能具有重要价值。基于酶联免疫吸附测定(ELISA)的血管扩张剂刺激磷蛋白(VASP)测定法相对于其他方法具有独特优势,且尚未在中国人群中使用。我们招募了10名健康志愿者和54例急性冠状动脉综合征患者。志愿者未接受治疗,患者接受氯吡格雷或替格瑞洛负荷剂量治疗。在基线和负荷剂量后8小时,使用流式细胞术(FC)-VASP和ELISA-VASP测量血小板反应性指数(PRI)。健康志愿者以及接受氯吡格雷或替格瑞洛治疗患者的血样在初始激活后冷冻保存1、2和4周。所有冷冻样品均使用ELISA-VASP进行检测。通过FC-VASP和ELISA-VASP评估的PRI相关性良好,在识别治疗中血小板反应性高或低方面显示出高度一致性。与未冷冻样品相比,在长达4周的冷冻样品中未观察到PRI结果有明显的时间依赖性变化。替格瑞洛治疗患者的PRI低于氯吡格雷治疗患者。ELISA-VASP能有效评估PRI,并且从冷冻标本获得的结果在检测前不受储存和运输的影响。在降低PRI方面,替格瑞洛优于氯吡格雷。

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