Preanalytic variables of thrombin generation: towards a standard procedure and validation of the method
Preanalytic variables of thrombin generation: towards a standard procedure and validation of the method
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DOI：
10.1111/jth.12012
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发表时间：
2012-12-01
影响因子：
10.4
通讯作者：
Spronk, H. M. H.
中科院分区：
文献类型：
--
作者：
Loeffen, R.;Kleinegris, M. -C. F.;Spronk, H. M. H.
.背景资料：凝血酶生成试验是评估血浆整体凝血潜力的敏感方法，但是，尽管它们在血栓形成研究中常用，但缺乏分析前条件的标准化。为了建立一个标准化的协议，我们分析了不同的预分析变量和验证校准的自动血栓图方法。方法和结果：用0、1和5 μ m组织因子（TF）评估凝血酶生成。凝血酶生成的变化主要归因于收集管的类型，主要是因为接触活化的变化。采集管还确定了其他分析前变量对凝血酶生成的影响，例如，对废弃管的需求、全血的储存和离心方法。关于采集系统，通过静脉导管或蝶形针抽取的血液显示出比使用传统针头进行静脉穿刺获得的血液显著更多的溶血。结果表明，凝血酶生成测量仍然需要废弃管。血液采集后，全血最好立即离心，以防止凝血蛋白的活化或降解，建议以10 000 x g进行第二次离心。解冻后，最好立即分析血浆，因为储存导致凝血酶生成结果超出参考样品的10%范围。在这些结果的基础上，我们建立了一个内部标准化方案，用于验证，导致1和5 pm TF触发的所有衍生参数的变异系数< 15%。结论：凝血酶生成受分析前条件的影响很大，表明需要一个国际标准化的协议。
. Background: Thrombin generation assays are sensitive methods for assessment of the overall clotting potential of plasma, but, despite their common use in thrombosis research, standardization of preanalytic conditions is lacking. In order to set up a standardized protocol, we analyzed different preanalytic variables and validated the calibrated automated thrombogram method. Methods and Results: Thrombin generation was assessed with 0, 1 and 5 pm tissue factor (TF). Variations in thrombin generation were mostly attributable to the type of collection tube, mainly because of variations in contact activation. The collection tube also determined the influence of other preanalytic variables on thrombin generation, e.g. the need for a discard tube, the storage of whole blood, and the centrifugation method. Regarding the collection system, blood drawn through intravenous catheters or butterfly needles showed significantly more hemolysis than blood obtained with venipuncture using conventional needles. The results showed that a discard tube is still needed for thrombin generation measurements. After blood collection, whole blood is best centrifuged immediately, to prevent activation or degradation of coagulation proteins, and a second centrifugation step at 10 000 x g is recommended. After thawing, plasma is best analyzed immediately, as storage resulted in thrombin generation results outside the 10% range of the reference sample. On the basis of these results, we set up an in-house standardized protocol, which was used for validation, resulting in coefficients of variations of < 15% for all derived parameters with both the 1 and 5 pm TF triggers. Conclusion: Thrombin generation was greatly influenced by preanalytic conditions, demonstrating the need for an international standardized protocol.