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首页> 外文期刊>Platelets >Optical multichannel (optimul) platelet aggregometry in 96-well plates as an additional method of platelet reactivity testing.
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Optical multichannel (optimul) platelet aggregometry in 96-well plates as an additional method of platelet reactivity testing.

机译：在96孔板上进行光学多通道（最佳）血小板凝集测定，作为血小板反应性测试的另一种方法。

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Platelet reactivity testing is important for the diagnosis of bleeding disorders, and increasingly to optimise anti-platelet therapy. Traditional light transmission aggregometry is considered the gold standard, whilst 96-well plate aggregometry, founded on similar principles, provides a higher throughput screening method. Despite the widespread use of both, methodologies and outputs vary widely between laboratories. We report a methodological approach towards providing a standardised optical detection of platelet aggregation (optimul method) based upon 96-well plate aggregometry. Individual wells of half-area 96-well plates were coated with gelatine and one of seven concentrations of arachidonic acid (AA), adenosine diphosphate (ADP), collagen, epinephrine (EPI), ristocetin, TRAP-6 amide or U46619, before being lyophilised, vacuum-sealed, foil-packed and stored at room temperature for up to 24 weeks. For platelet testing, 40 microl of platelet-rich plasma was added to each well. Platelet aggregation was determined by changes in light absorbance, release of ATP/ADP by luminescence and release of thromboxane (TX) A(2) by ELISA. Some experiments were conducted in the presence of aspirin (30 microM) or prasugrel active metabolite (PAM; 3 microM). Optimul plates stored for up to 12 weeks permitted reliable detection of concentration-dependent platelet aggregation, ATP/ADP release and TXA production. PAM caused reductions in platelet responses to AA, ADP, collagen, EPI, TRAP-6 and U46619, whilst aspirin inhibited responses to AA, collagen and EPI. We conclude that the optimul method offers a viable, standardised approach, allowing platelet reactivity testing and could provide a broad platelet function analysis without the need for dedicated equipment.
机译：血小板反应性测试对出血性疾病的诊断非常重要，并且越来越多地用于优化抗血小板治疗。传统的透光聚集法被认为是金标准，而基于相似原理的96孔板聚集法则提供了更高的通量筛选方法。尽管两者均被广泛使用，但实验室之间的方法和产出差异很大。我们报告了一种方法学方法，旨在基于96孔板凝集法提供标准化的血小板凝集光学检测（最佳方法）。在半区域96孔板的各个孔中涂明胶，然后涂七种浓度的花生四烯酸（AA），二磷酸腺苷（ADP），胶原蛋白，肾上腺素（EPI），瑞斯托菌素，TRAP-6酰胺或U46619中的一种。冻干，真空密封，箔包装并在室温下保存长达24周。为了进行血小板测试，将40微升富含血小板的血浆添加到每个孔中。通过光吸收的变化，通过发光释放ATP / ADP和通过ELISA释放血栓烷（TX）A（2）来确定血小板聚集。在阿司匹林（30 microM）或普拉格雷活性代谢物（PAM; 3 microM）的存在下进行了一些实验。最佳平板可保存长达12周，从而能够可靠地检测出浓度依赖性血小板聚集，ATP / ADP释放和TXA产生。 PAM导致血小板对AA，ADP，胶原蛋白，EPI，TRAP-6和U46619的反应减少，而阿司匹林抑制对AA，胶原蛋白和EPI的反应。我们得出结论，最佳方法提供了一种可行的，标准化的方法，可以进行血小板反应性测试，并且可以在不需要专用设备的情况下进行广泛的血小板功能分析。

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