首页|基于VASP评估氯吡格雷抗血小板疗效方法的建立及应用

基于VASP评估氯吡格雷抗血小板疗效方法的建立及应用

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目的 建立直接化学发光免疫分析法检测血管扩张刺激磷蛋白(vasodilator stimulating phos-phoprotein，VASP)以评估氯吡格雷的抗血小板效果。方法 以超顺磁微粒为固相，吖啶酯为标记物，基于直接化学发光免疫分析技术建立新的检测血管扩张刺激磷蛋白磷酸化的方法，评估空白限、线性、批内精密度、回收率、热稳定性性能;同时测试全血样本，与PLT VASP/P2Y12流式检测试剂盒测定结果进行比对。结果 新方法的空白限小于0。5 ng·mL-1，线性范围为0。06～50。53 ng·mL-1，批内精密度RSD小于5。0％，回收率大于90％，热稳定性结果良好，45 min完成检测，与PLT VASP/P2Y12流式检测试剂盒测定结果一致性大于95％。结论 基于直接化学发光免疫分析技术建立新的检测血管扩张刺激磷蛋白磷酸化的方法性能优异、检测时间短，与上市产品结果一致，能够给氯吡格雷临床抗血小板个体化用药提供参考。
Establishment of a new method for evaluating the antiplatelet effect of clopidogrel based on the detection of vasodilator stimulating phosphoprotein by direct chemiluminescence immunoassay
Objective To establish a direct chemiluminescence immunoassay to detect vasodilator stimulating phosphoprotein(VASP)for the evaluation of the antiplatelet effect of clopidogrel.Methods Using superparamagnetic particles as the solid phase and acridine ester as the marker,a new method for detecting the phosphorylation of phosphoprotein stimulated by vasodilation was established based on the direct chemiluminescence immunoassay technology,and the limit of blank,linearity,intra-day precision,recovery rate and thermal stability were evaluated.Concurrently,the results of the whole blood sample determination were compared with the PLT VASP/P2Y12 flow detection kit of Stago.Results The limit of blank was less than 0.5 ng·mL-1,the linear range was 0.06-50.53 ng·mL-1,the coefficient of variation for intra-day precision was less than 5.0％,the recovery rate was more than 90％and the results of thermal stability were good.The test was completed in 45 minutes,and the consistency of the measured results with PLT VASP/P2Y12 flow detection kit of Stago France was greater than 95％.Conclusion A new method based on direct chemiluminescence immunoassay for the detection of vasodilation-stimulated phosphoprotein phosphorylation had excellent performance,short detection time,and good consistency with the results of marketed products,which can provide effective data reference for clopidogrel clinical antiplatelet drugs.

direct chemiluminescence immunoassayvasodilator stimulating phosphoproteinclopidogrelantiplatelet effect

李明勇、石雅宁、王晖、覃丽

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湖南中医药大学 药学院 干细胞中药调控与应用实验室,湖南长沙 410208

湖南远璟生物技术有限公司,湖南长沙 410201

中南大学湘雅医学院,湖南长沙 410031

直接化学发光免疫法 血管扩张刺激磷蛋白 氯吡格雷 抗血小板效果

国家自然科学基金面上项目湖南省自然科学基金科药联合基金湖南省教育厅项目湖南省教育厅项目湖南省卫生健康委重点指导课题长沙市科技局科研项目中药粉体与创新药物省部共建国家重点实验室培育基地开放基金

81973668CX2022079220A37522B0355202213055529kq200406021PTKF1004

2024

10.14066/j.cnki.cn21-1349/r.2023.0593

沈阳药科大学学报
沈阳药科大学

沈阳药科大学学报

CSTPCD
影响因子：0.604
ISSN：1006-2858
年,卷(期)：2024.41(4)
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