Department of Laboratory Medicine, Keio University School of Medicine, Tokyo, Japan.
Clinical Laboratory, Keio University Hospital, Tokyo, Japan.
Clin Chim Acta. 2024 Aug 15;562:119887. doi: 10.1016/j.cca.2024.119887. Epub 2024 Jul 24.
Clot waveform analysis (CWA) reportedly enhances the interpretation of clotting time measurement. This study aimed to compare CWA between prothrombin time (PT) and activated partial thromboplastin time (APTT) assays for better understanding how to apply CWA for assessing effects of direct oral anticoagulants (DOACs).
Samples were prepared by spiking plasma with rivaroxaban, apixaban, edoxaban, or dabigatran. To compensate the influence of fibrinogen, CWA parameters were adjusted by unifying maximum changes in transmittance in clotting reaction curves detected by the optical system.
Non-adjusted PT-CWA parameters unexpectedly rose at low drug concentrations but declined at high drug concentrations while adjusted PT-CWA parameters exhibited dose-dependent decrease. Both non-adjusted and adjusted APTT-CWA parameters showed dose-dependent decrease. Adjusted CWA parameters were applicable to Hill plot analysis. All DOACs exhibited Hill coefficients indicating positively cooperative effects regarding most adjusted PT-CWA parameters. Regarding adjusted APTT-CWA parameters, rivaroxaban, apixaban, and edoxaban exhibited Hill coefficients indicating no or negatively cooperative effects. The observed differences between PT-CWA and APTT-CWA suggested the implication of thrombin positive feedback in DOAC effects.
The results revealed distinct features of DOAC effects in extrinsic and intrinsic pathways. To ascertain the clinical implication, further studies using clinical samples are needed.
据报道,凝块波形分析(CWA)可增强对凝血时间测量的解释。本研究旨在比较凝血酶原时间(PT)和活化部分凝血活酶时间(APTT)测定中的 CWA,以更好地了解如何应用 CWA 评估直接口服抗凝剂(DOAC)的作用。
通过向血浆中添加利伐沙班、阿哌沙班、依度沙班或达比加群来制备样品。为了补偿纤维蛋白原的影响,通过统一光学系统检测到的凝血反应曲线中透光率的最大变化来调整 CWA 参数。
未经调整的 PT-CWA 参数在低药物浓度下意外升高,但在高药物浓度下下降,而调整后的 PT-CWA 参数则呈剂量依赖性下降。未经调整和调整后的 APTT-CWA 参数均呈剂量依赖性下降。调整后的 CWA 参数适用于 Hill 图分析。所有 DOAC 均表现出表明对大多数调整后的 PT-CWA 参数具有正协同作用的 Hill 系数。关于调整后的 APTT-CWA 参数,利伐沙班、阿哌沙班和依度沙班表现出表明无协同作用或负协同作用的 Hill 系数。PT-CWA 和 APTT-CWA 之间观察到的差异表明 DOAC 作用中存在凝血酶正反馈。
研究结果揭示了 DOAC 在外源性和内源性途径中的作用的独特特征。为了确定临床意义,需要使用临床样本进行进一步研究。