Nieuwenhuizen W, Hoegee-De Nobel E, Laterveer R
IVVO-TNO, Gaubius Laboratory, Leiden, The Netherlands.
Thromb Haemost. 1992 Sep 7;68(3):273-7.
Soluble fibrin is considered as a molecular marker for intravascular fibrin formation, and impending thrombotic events. Most of the existing assays are less suitable for routine clinical applications and their specificity may be limited. We have developed a sandwich-type EIA with a fibrin-specific MoAb described by us before (Proc Natl Acad Sci 1989; 86: 8951) as the capture antibody. An other MoAb (G8; Thromb Haemostas 1988; 60: 145) with an epitope in the carboxyl-terminal sections of the fibrin alpha-chains, was labeled with peroxidase and used as the tagging antibody. The EIA is calibrated against plasma spiked with known concentrations of soluble fibrin. The time-to-result of the EIA is only 1.5 h. Concentrations as low as 0.5 micrograms soluble fibrin/ml plasma are readily measureable. Heparin has no effect on the results. Fibrinogen and fibrin(ogen) degradation products are not detected. The values of fibrinopeptide A and soluble fibrin values found with the "COA-SET soluble fibrin" assay correlated well with the soluble fibrin values found with our EIA i.e. r = 0.998 and 0.984, respectively. The run-to-run variabilities were 7.9% and 6.6% for samples with low and high soluble fibrin concentrations, respectively. The within-run variabilities were 2.5, 1.8, 4.0 and 4.6% for samples with 1, 0.5, 0.25 and 0.125 micrograms soluble fibrin/ml, respectively. The sensitivity, specificity, accuracy and short time-to-result make our EIA suitable for routine clinical applications and the monitoring of the effectivity of heparinization.
可溶性纤维蛋白被视为血管内纤维蛋白形成和即将发生的血栓事件的分子标志物。现有的大多数检测方法不太适合常规临床应用，其特异性可能有限。我们开发了一种夹心型酶免疫分析方法，使用我们之前描述的（《美国国家科学院院刊》1989年；86:8951）纤维蛋白特异性单克隆抗体作为捕获抗体。另一种单克隆抗体（G8；《血栓与止血》1988年；60:145），其表位位于纤维蛋白α链的羧基末端部分，用辣根过氧化物酶标记并用作标记抗体。该酶免疫分析方法以添加已知浓度可溶性纤维蛋白的血浆进行校准。酶免疫分析方法的出结果时间仅为1.5小时。血浆中低至0.5微克可溶性纤维蛋白/毫升的浓度很容易测量。肝素对结果无影响。未检测到纤维蛋白原和纤维蛋白（原）降解产物。用“COA-SET可溶性纤维蛋白”检测方法测得的纤维蛋白肽A值和可溶性纤维蛋白值与我们的酶免疫分析方法测得的可溶性纤维蛋白值相关性良好，即r分别为0.998和0.984。低可溶性纤维蛋白浓度和高可溶性纤维蛋白浓度样本的批间变异系数分别为7.9%和6.6%。对于可溶性纤维蛋白浓度分别为1、0.5、0.25和0.125微克/毫升的样本，批内变异系数分别为2.5%、1.8%、4.0%和4.6%。该酶免疫分析方法的灵敏度、特异性、准确性和较短的出结果时间使其适用于常规临床应用以及监测肝素化的有效性。
