Determination of Soluble Fibrin in Plasma by a Rapid and Quantitative Spectrophotometric Assay
Determination of Soluble Fibrin in Plasma by a Rapid and Quantitative Spectrophotometric Assay
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通过快速定量分光光度测定法测定血浆中的可溶性纤维蛋白
DOI：
10.1055/s-0038-1661519
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发表时间：
1986
影响因子：
6.7
通讯作者：
M. Rånby
中科院分区：
文献类型：
--
作者：
B. Wiman;M. Rånby
本文建立了一种快速、灵敏、定量测定血浆中可溶性纤维蛋白的分光光度法。该方法基于纤维蛋白刺激组织型纤溶酶原激活剂（t-PA）激活纤溶酶原的原理。将含有纤维蛋白的样品与t-PA、纤溶酶原和纤溶酶敏感的显色底物一起孵育。吸光度的增加取决于样品中的纤维蛋白浓度。在血浆样品中，由于存在α 2-抗纤溶酶，观察到初始滞后期。然而，发现在反应后期A405每平方分钟的变化与纤维蛋白含量成比例，并在原点产生线性标准曲线截距。血纤蛋白原的检测限约为5 nmol/L。在20和75 nmol/L水平下，健康个体血浆样品的回收率实验结果非常好。“运行内”变异（CV）确定为3.7%。在血浆中加入微量凝血酶后，纤维蛋白的形成可用该方法监测。8例健康人血浆中纤维蛋白含量为9.2 ± 1.9nmol/L。46例疑似止血障碍患者的血浆样本中可溶性纤维蛋白水平较高（53 ± 62 nmol/L）。46份样品中有7份的浓度高于150 nmol/L。
Summary A rapid, sensitive and quantitative spectrophotometric assay of soluble fibrin in plasma samples has been developed. The method is based on the principle that fibrin stimulates the activation of plasminogen by tissue plasminogen activator (t-PA). A sample containing fibrin is incubated with t-PA, plasminogen and a plasmin-sensitive chromogenic substrate. An increase in absorbance which is dependent on the fibrin concentration in the sample is obtained. In plasma samples an initial lag-phase, due to the presence of alpha2-antiplasmin is observed. However, the change in A405 per square minute during the later part of the reaction was found to be proportional to the fibrin content and to generate linear standard curve intercepts at the origin. The detection limit of bathroxobin-digested fibrinogen added to plasma was about 5 nmol/L. Recovery experiments at 20 and 75 nmol/L levels in plasma samples from healthy individuals were excellent. The “within-run” variation (CV) was determined as 3.7%. The formation of fibrin after addition of minute amounts of thrombin to plasma could be monitored with the method. Plasma from 8 healthy individuals was found to contain about 9.2 ± 1.9 nmol/L fibrin. Plasma samples from 46 patients with a suspected haemostatic disturbance had higher levels of soluble fibrin (53 ± 62 nmol/L). Seven of the 46 samples had concentrations above 150 nmol/L.