Benson R E, Catalfamo J L, Brooks M, Dodds W J
Wadsworth Center for Laboratories & Research, New York State Department of Health, Albany 12201-0509.
J Immunoassay. 1991;12(3):371-90. doi: 10.1080/01971529108055078.
We have developed an ELISA specific for canine von Willebrand factor antigen (vWF:Ag) that also strongly reacts with the VWF:Ag of humans and many other vertebrates. This assay was designed to avoid the use of immunoreagents of human origin, however, commercially available antibodies to human vWF:Ag may also be used. von Willebrand factor (vWF) was quantitated using a modified double-sandwich ELISA with polyclonal antibodies specific for canine vWF:Ag. The assay was as sensitive for measuring canine vWF:Ag as previously published immuno-radiometric assays and the most sensitive ELISA for human vWF:Ag. Employing commercially available antibodies to human vWF:Ag in the same double-sandwich configuration, the lower limit of detection for human vWF:Ag was 4.8 x 10(-6) units/ml, lower by a factor of ten than previously reported ELISAs. In addition, a wide range of vWF:Ag levels can be determined with just a single plasma dilution. The assay readily distinguishes type III von Willebrand disease from other types of von Willebrand disease having very low levels of vWF. This vWF ELISA can be used to evaluate large numbers of plasma samples simultaneously and is therefore well-suited for large-scale screening programs.
我们开发了一种针对犬血管性血友病因子抗原（vWF:Ag）的酶联免疫吸附测定（ELISA），该测定也能与人类及许多其他脊椎动物的VWF:Ag发生强烈反应。此检测方法旨在避免使用源自人类的免疫试剂，不过也可使用市售的抗人类vWF:Ag抗体。使用对犬vWF:Ag具有特异性的多克隆抗体，通过改良的双夹心ELISA对血管性血友病因子（vWF）进行定量。该检测方法在测量犬vWF:Ag时的灵敏度与先前发表的免疫放射测定法相同，且是检测人类vWF:Ag最灵敏的ELISA方法。采用市售的抗人类vWF:Ag抗体以相同的双夹心形式进行检测，人类vWF:Ag的检测下限为4.8×10⁻⁶单位/毫升，比先前报道的ELISA低一个数量级。此外，仅用单一血浆稀释度就能测定广泛范围的vWF:Ag水平。该检测方法能轻松区分III型血管性血友病与其他vWF水平极低的血管性血友病类型。这种vWF ELISA可同时用于评估大量血浆样本，因此非常适合大规模筛查项目。
