Letellier C, Delangre A, De Smet A, Kerkhofs P
Veterinary and Agrochemical Research Centre, 1180 Brussels, Belgium.
Vet Microbiol. 2001 Dec 4;83(4):301-15. doi: 10.1016/s0378-1135(01)00430-8.
A panel of seven monoclonal antibodies (MAbs) directed against the bovine herpesvirus-1 (BHV-1) glycoprotein E (gE) was obtained. For that purpose, mice were either tolerized to BHV-1 gE-negative virus and then immunized with wild type BHV-1 or immunized with plasmid DNA expressing the gE and gI glycoproteins. The MAbs were characterized by their reactivity with the gE protein or the gE/gI complex and by competition experiments. Results showed that the MAbs were directed against three antigenic domains, two located on the gE glycoprotein and one on the gE/gI complex. Blocking experiments were performed with sera from experimentally vaccinated and infected cattle. A competition was observed between gE-positive bovine sera and six of the seven MAbs. The bovine sera thus recognized two of the three antigenic sites. Field sera were then tested in blocking enzyme-linked immunosorbent assay using one horseradish peroxidase-conjugated MAb. A specificity of 98.2% and a sensitivity of 98.2% compared to the commercially available test were observed.
获得了一组针对牛疱疹病毒1型（BHV-1）糖蛋白E（gE）的七种单克隆抗体（MAb）。为此，小鼠要么用BHV-1 gE阴性病毒进行耐受处理，然后用野生型BHV-1免疫，要么用表达gE和gI糖蛋白的质粒DNA免疫。通过它们与gE蛋白或gE/gI复合物的反应性以及竞争实验对这些单克隆抗体进行了表征。结果表明，这些单克隆抗体针对三个抗原结构域，其中两个位于gE糖蛋白上，一个位于gE/gI复合物上。用来自经实验性接种和感染牛的血清进行了阻断实验。在gE阳性牛血清与七种单克隆抗体中的六种之间观察到了竞争。因此，牛血清识别出了三个抗原位点中的两个。然后使用一种辣根过氧化物酶偶联的单克隆抗体，在阻断酶联免疫吸附试验中对现场血清进行了检测。与市售检测方法相比，观察到特异性为98.2%，灵敏度为98.2%。
