Kalina Uwe, Stöhr Hans-Arnold, Bickhard Heike, Knaub Sigurd, Siboni Simona M, Mannucci Pier M, Peyvandi Flora
Department of Clinical Research and Development, CSL Behring GmbH, Marburg, Germany.
Blood Coagul Fibrinolysis. 2008 Dec;19(8):777-83. doi: 10.1097/MBC.0b013e32830ef90c.
To characterize a functional assay for circulating fibrinogen based on rotational thrombelastography. Maximum clot firmness was determined by rotational thrombelastography in normal human plasma pool, fibrinogen-deficient plasma pool, normal whole blood, and individual plasma samples from 17 patients with fibrinogen deficiency. Plasma samples spiked with varying concentrations of exogenous fibrinogen were also measured. Results were compared with enzyme-linked immunosorbent assay and Clauss assay. The impact of sample freezing and filtration and use of cytochalasin D were also investigated. Over the tested range of 0-3 mg/ml added exogenous fibrinogen, the maximum clot firmness standard curve for determination of fibrinogen in plasma pools (n = 7) was linear (r2 = 0.97). Maximum clot firmness was highly linearly correlated both with Clauss assay (r2 = 0.93) and enzyme-linked immunosorbent assay (r2 = 0.95). In unspiked plasma samples from individual patients with fibrinogen deficiency, fibrinogen was undetectable by rotational thromboelastography. By all evaluated methods, the response to spiking with fibrinogen in such samples coincided closely in patients with afibrinogenemia and hypofibrinogenemia. In dysfibrinogenemia, smaller Clauss assay responses to spiking were observed, whereas the enzyme-linked immunosorbent assay response was variable. Maximum clot firmness was the only evaluated method of fibrinogen assessment to yield consistent results across all categories of fibrinogen deficiency. These in-vitro results suggest the potential clinical utility of rotational thromboelastography as a versatile method for monitoring the response to fibrinogen concentrate among patients with fibrinogen deficiency. Clinical investigations using rotational thromboelastography after in-vivo fibrinogen administration to patients with congenital fibrinogen deficiency are warranted.
基于旋转血栓弹力图法对循环纤维蛋白原功能测定进行特性描述。通过旋转血栓弹力图法测定正常人血浆池、纤维蛋白原缺乏血浆池、正常全血以及17例纤维蛋白原缺乏患者的个体血浆样本中的最大血凝块硬度。还对添加不同浓度外源性纤维蛋白原的血浆样本进行了检测。将结果与酶联免疫吸附测定法和Clauss测定法进行比较。同时研究了样本冷冻、过滤以及细胞松弛素D的使用所产生的影响。在添加0 - 3mg/ml外源性纤维蛋白原的测试范围内，血浆池（n = 7）中用于测定纤维蛋白原的最大血凝块硬度标准曲线呈线性（r2 = 0.97）。最大血凝块硬度与Clauss测定法（r2 = 0.93）和酶联免疫吸附测定法（r2 = 0.95）均高度线性相关。在纤维蛋白原缺乏个体患者的未添加样本中，旋转血栓弹力图法检测不到纤维蛋白原。通过所有评估方法，在无纤维蛋白原血症和低纤维蛋白原血症患者中，此类样本对添加纤维蛋白原的反应密切吻合。在异常纤维蛋白原血症中，观察到Clauss测定法对添加的反应较小，而酶联免疫吸附测定法的反应则有所不同。最大血凝块硬度是评估纤维蛋白原的唯一方法，在所有类型的纤维蛋白原缺乏中均能产生一致的结果。这些体外结果表明，旋转血栓弹力图法作为一种通用方法监测纤维蛋白原缺乏患者对纤维蛋白原浓缩物的反应具有潜在临床应用价值。有必要对先天性纤维蛋白原缺乏患者进行体内给予纤维蛋白原后使用旋转血栓弹力图法的临床研究。
