Favaloro E J, Henniker A, Facey D, Hertzberg M
Department of Haematology, Institute of Clinical Pathology and Medical Research, Westmead Hospital, NSW, Australia.
Thromb Haemost. 2000 Oct;84(4):541-7.
Discrimination of von Willebrand's Disease (VWD) subtypes is important since it influences management. Qualitative [ie Type 2A, 2B, 2M] defects exhibit von Willebrand factor (VWF) discordance and give high VWF:Ag to VWF:'activity' ratios. Classically, VWF:'activity' is assessed using the VWF:RCof assay. The VWF:CBA is an ELISA-based VWF-functional adhesive assay which has consistently proved to be superior to VWF:RCof. A commercially available monoclonal antibody (MAB) based ELISA assay system claimed to mimic a VWF:RCof-like activity has also been recently described ('SE'), as has the production and characterisation of a large number [n = 10] of locally generated anti-VWF MAB. In the current study, we have adapted these MAB to in-house ELISA assays to assess their utility for VWD diagnosis and subtype discrimination, and to compare them with other assay systems. Thus, the VWF:CBA, VWF:RCof by agglutination, the SE assay, and in-house MAB based assays have been directly compared for their ability to discriminate Type 1 [n = 9] from Type 2 VWD samples [phenotypes 2A and 2B; n = 11]. In summary, MAB-based systems can be used to measure VWF and confirm a diagnosis of VWD, as well as exhibiting some VWD-subtype-discriminatory capabilities. However, better evidence of VWF-discordance was usually achieved using the VWF:RCof (agglutination) assay, while the greatest degree of VWF-discordance was consistently observed using the VWF:CBA assay. In conclusion, the VWF:CBA assay proved to offer the best diagnostic predictive tool for a Type 2 VWD defect, while MAB-based systems appear to be less effective in this regard.
鉴别血管性血友病(VWD)的亚型很重要,因为这会影响治疗管理。定性缺陷(即2A型、2B型、2M型)表现为血管性血友病因子(VWF)不一致,且VWF:抗原与VWF:活性的比值较高。传统上,VWF:活性通过VWF:瑞斯托霉素辅因子(VWF:RCof)测定法进行评估。VWF:胶原结合测定法(VWF:CBA)是一种基于酶联免疫吸附测定(ELISA)的VWF功能黏附测定法,一直被证明优于VWF:RCof测定法。最近还描述了一种基于市售单克隆抗体(MAB)的ELISA测定系统,声称可模拟VWF:RCof样活性(“SE”),以及大量(n = 10)本地产生的抗VWF MAB的制备和表征。在本研究中,我们将这些MAB应用于内部ELISA测定,以评估其在VWD诊断和亚型鉴别中的效用,并与其他测定系统进行比较。因此,我们直接比较了VWF:CBA、凝集法VWF:RCof、SE测定法和基于内部MAB的测定法鉴别1型VWD样本(n = 9)与2型VWD样本(2A和2B型表型;n = 11)的能力。总之,基于MAB的系统可用于测量VWF并确诊VWD,也具有一定的VWD亚型鉴别能力。然而,通常使用VWF:RCof(凝集)测定法能获得更好的VWF不一致证据,而使用VWF:CBA测定法始终能观察到最大程度的VWF不一致。总之,VWF:CBA测定法被证明是诊断2型VWD缺陷的最佳预测工具,而基于MAB的系统在这方面似乎效果较差。