DEVELOPMENT OF A SIMPLE COLLAGEN BASED ELISA ASSAY AIDS IN THE DIAGNOSIS OF, AND PERMITS SENSITIVE DISCRIMINATION BETWEEN TYPE-I AND TYPE-II, VONWILLEBRANDS DISEASE
DEVELOPMENT OF A SIMPLE COLLAGEN BASED ELISA ASSAY AIDS IN THE DIAGNOSIS OF, AND PERMITS SENSITIVE DISCRIMINATION BETWEEN TYPE-I AND TYPE-II, VONWILLEBRANDS DISEASE
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DOI：
10.1097/00001721-199104000-00011
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发表时间：
1991-04-01
影响因子：
1.1
通讯作者：
KOUTTS, J
中科院分区：
文献类型：
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作者：
FAVALORO, EJ;GRISPO, L;KOUTTS, J
我们开发并评估了一种基于elisa的胶原结合试验（CBA）作为血管性血友病（vWD）诊断和分类的辅助方法。该试验操作简单，似乎能够区分II型vWD和I型vWD。使用来自受影响和未受影响患者或正常人的血浆样本，使用CBA获得的数据直接与同时从标准血管性血友病因子抗原（蛋白；vWFAg） ELISA和标准利斯托司汀辅助因子（rof）测定中获得的数据进行比较。vWD患者（ⅰ型和ⅱ型）血浆中vWF水平总体降低。I型患者的vWFAg、CBA、RCof的平均水平分别为47.3、60.7、31.1 (n = 37)， II型患者的平均水平为34.9、1.6、11.9 （n = 16）。然而，尽管在I型vWD患者中检测到vWF水平降低，但CBA和vWFAg的vWF结合值几乎具有可比性（即vWFAg:CBA的比值通常小于或等于1.0）。因此，这些比值值与使用个体正常供体或非vwd患者血浆所观察到的值相似。另一方面，II型vWD患者的血浆显示出明显不同的值，vWFAg:CBA比值升高（> 8.0）与这些患者几乎没有CBA结合相一致。因此，本文报道的CBA似乎确实构成了一种新的功能测定方法，能够检测受影响的vWD患者血浆中vWF的定性差异。它的简单性使其成为目前研究vWF或vWD患者的任何实验室的理想检测方法，并可能允许未来在受影响的患者中发现新的功能失调形式的vWF。
We have developed and evaluated an ELISA-based collagen binding assay (CBA) as an aid in the diagnosis and classification of von Willebrand's disease (vWD). The assay is simple to perform, and appears capable of differentiating Type II vWD from Type I vWD. Using plasma samples from both affected and non-affected patients, or from normal individuals, data obtained using the CBA were directly compared to data simultaneously derived from a standard von Willebrand factor antigen (protein; vWFAg) ELISA, and from a standard ristocetin cofactor (RCof) assay. Plasma derived from vWD patients (both Type I and Type II) showed overall reduced levels of vWF as detected by all three assays. Mean levels as a per cent of normal for vWFAg, CBA, RCof were 47.3, 60.7, 31.1 for Type I patients (n = 37), and 34.9, 1.6, 11.9 for Type II patients (n = 16) respectively. However despite the reduced vWF levels detected in Type I vWD patients, vWF binding values for both the CBA and vWFAg showed near comparability (i.e. vWFAg:CBA ratio generally less-than-or-equal-to 1.0). These ratio values were thus similar to those observed using plasma derived from either individual normal donors, or from non-vWD affected patients. On the other hand, plasma from Type II vWD affected patients showed markedly disparent values, with increased (> 8.0) vWFAg:CBA ratios coincident with virtually absent CBA binding in these patients. Thus, the CBA as reported here does appear to constitute a novel functional assay capable of detecting qualitative vWF differences in plasma of affected vWD patients. Its simplicity makes it an ideal assay for any laboratory current investigating vWF or vWD patients, and may allow the future discovery of new dysfunctional forms of vWF in affected patients.