Lattuada Antonella, Preda Luigi, Sacchi Elisabetta, Gallo Luciana, Federici Augusto B, Rossi Edoardo
Immunohematology and Blood Transfusion Service, 'L. Sacco' Hospital, Milan
Blood Coagul Fibrinolysis. 2004 Sep;15(6):505-11. doi: 10.1097/00001721-200408000-00011.
We have set up a rapid assay for measuring the activity of the von Willebrand factor ristocetin cofactor (VWF : RCo) using an automated coagulometer (ACL 9000; Instrumentation Laboratory, Lexington, Massachusetts, USA) and commercially available lyophilized platelet reagents (Dade-Behring, Marburg, Germany). Since VWF : RCo tested with the coagulometer (ACL-VWF : RCo) did not require loading, calculation of von Willebrand factor (VWF) activity or pretreatment of samples (calibration standard and tested plasmas), we thought it might be useful for rapid, automatic screening of VWF activity. To assess the precision and the potency of this ACL-VWF : RCo, we tested the assay in this laboratory's internal normal and abnormal controls, in a group of 67 healthy individuals and in 28 patients with different types of von Willebrand disease (VWD). We compared the ACL-VWF : RCo findings with those of the standard agglutination test (Agg-VWF : RCo), normalizing both assays against VWF antigen (VWF : Ag) measured by 'automatic' and standard enzyme-linked immunosorbent assay 'in-house' methods, to calculate the VWF : RCo/VWF Ag ratios. The within-assay and between-assay repeatability of the automatic ACL-VWF : RCo gave coefficients of variation < 5%, and reproducibility in normal and abnormal laboratory controls gave coefficients of variation of 7.9 and 9.3%, respectively. In VWD patients the results were equivalent to those of the standard Agg-VWF : RCo assay but, because of the good sensitivity, the ACL-VWF : RCo was more accurate in evaluating the VWF : RCo and establishing the VWF : RCo/VWF : Ag ratios in VWD patients with low VWF levels. Moreover, this ACL-VWF : RCo is faster than Agg-VWF : RCo, providing results of calibration curves and of 10 patient samples within 15 min. We can conclude that this automatic ACL-VWF : RCo gives a reliable and useful measure of VWF activity and can be proposed as a screening test for rapid diagnosis of VWD even in patients with low VWF levels in plasma.
我们使用自动凝血分析仪(ACL 9000;美国马萨诸塞州列克星敦市仪器实验室公司)和市售冻干血小板试剂(德国马尔堡达德 - 贝林公司)建立了一种快速检测血管性血友病因子瑞斯托霉素辅因子(VWF : RCo)活性的方法。由于用该凝血分析仪检测的VWF : RCo(ACL - VWF : RCo)无需加样、计算血管性血友病因子(VWF)活性或对样品(校准标准品和待测血浆)进行预处理,我们认为它可能有助于快速、自动筛查VWF活性。为评估这种ACL - VWF : RCo的精密度和效能,我们在本实验室内部的正常和异常对照品、67名健康个体以及28例不同类型血管性血友病(VWD)患者中对该检测方法进行了测试。我们将ACL - VWF : RCo的检测结果与标准凝集试验(Agg - VWF : RCo)的结果进行比较,将两种检测方法针对通过“自动”和标准酶联免疫吸附测定“内部”方法测得的VWF抗原(VWF : Ag)进行标准化,以计算VWF : RCo/VWF : Ag比值。自动ACL - VWF : RCo的批内和批间重复性变异系数<5%,在正常和异常实验室对照品中的重现性变异系数分别为7.9%和9.3%。在VWD患者中,结果与标准Agg - VWF : RCo检测结果相当,但由于灵敏度高,ACL - VWF : RCo在评估VWF水平较低的VWD患者的VWF : RCo以及建立VWF : RCo/VWF : Ag比值方面更准确。此外,这种ACL - VWF : RCo比Agg - VWF : RCo更快,可在15分钟内提供校准曲线和10份患者样品的结果。我们可以得出结论,这种自动ACL - VWF : RCo能可靠且有效地检测VWF活性,甚至对于血浆中VWF水平较低的患者,也可作为快速诊断VWD的筛查试验。