Durham P J, Hassard L E
Department of Veterinary Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada.
Vet Microbiol. 1990 Mar;22(1):1-10. doi: 10.1016/0378-1135(90)90118-f.
A single dilution enzyme-linked immunosorbent assay (ELISA) was developed to detect antibodies to bovine viral diarrhea (BVD) virus in cattle sera. Viral antigen (NADL strain) was grown in a pig kidney cell line (PK15), and after removal of nuclear debris, was purified by ultracentrifugation through a potassium tartrate cushion. Antigen grown in embryonic bovine tracheal epithelial cells was also satisfactory. The test used a high salt buffer to minimize nonspecific reactivity, polyethylene glycol to enhance the reaction, and Protein G as the labelling agent. Comparative testing with the virus neutralization test (VNT) showed the ELISA results to have a high level of correlation with the VNT titers (r = 0.83). In vaccinated animals the ELISA detected antibodies earlier than the VNT. All animals sampled from a BVD-free herd were negative for BVD antibody. The single dilution test showed close agreement (r = 0.84) with ELISA values obtained using a serial dilution technique, and also proved to have a high level of reproducibility. The test procedures were relatively easy to carry out, and were economic in their use of materials.
开发了一种单稀释酶联免疫吸附测定（ELISA）法，用于检测牛血清中针对牛病毒性腹泻（BVD）病毒的抗体。病毒抗原（NADL株）在猪肾细胞系（PK15）中培养，去除核碎片后，通过酒石酸钾垫层超速离心进行纯化。在胚胎牛气管上皮细胞中培养的抗原也令人满意。该试验使用高盐缓冲液以尽量减少非特异性反应，用聚乙二醇增强反应，并使用蛋白G作为标记剂。与病毒中和试验（VNT）的对比检测表明，ELISA结果与VNT滴度具有高度相关性（r = 0.83）。在接种疫苗的动物中，ELISA比VNT更早检测到抗体。从无BVD牛群中采集的所有动物的BVD抗体均为阴性。单稀释试验与采用系列稀释技术获得的ELISA值显示出密切一致性（r = 0.84），并且还证明具有高度的可重复性。该试验程序相对易于实施，且材料使用经济。
