Department of Epidemiology and Vety. Preventive medicine, College of Veterinary Sciences, Veterinary University, Mathura (UP), India.
Asian Pac J Trop Med. 2011 May;4(5):363-6. doi: 10.1016/S1995-7645(11)60104-1. Epub 2011 Jun 22.
To develop a standard enzyme-linked immunosorbent assay (ELISA) for the detection of bovine herpesvirus type 1 (BHV-1).
The assay was based on hyperimmune rabbit and guinea pig antisera raised against purified BHV-1. Polyethylene glycol precipitation and sucrose density gradient methods were adopted for viral concentration and purification. Antisera were raised using Freund's adjuvant followed by extraction of IgG of high purity.
Optimum antisera dilutions as determined by titrations were chosen as 1:4 000, whereas the conjugate was used at 1:2 000 dilution. Using 95 clinical specimens, the ELISA test showed a sensitivity and specificity of 91.90 % and 93.10 %, respectively when compared to PCR. The cut-off value was fixed at 0.15 (A(490)) and a P/N ratio of >1.30 indicated a significant positive reaction.
The results have demonstrated that this ELISA could efficiently detect BHV-1 and can be used as an important diagnostic tool.
开发一种用于检测牛疱疹病毒 1 型（BHV-1）的标准酶联免疫吸附试验（ELISA）。
该检测基于针对纯化的 BHV-1 产生的高免兔和豚鼠抗血清。采用聚乙二醇沉淀和蔗糖密度梯度法进行病毒浓缩和纯化。使用弗氏佐剂进行抗血清的制备，随后提取高纯度的 IgG。
通过滴定确定的最佳抗血清稀释度为 1:4000，而结合物的稀释度为 1:2000。使用 95 份临床标本与 PCR 相比，ELISA 检测的敏感性和特异性分别为 91.90%和 93.10%。将截断值固定在 0.15（A(490)），P/N 比值>1.30 表示显著阳性反应。
结果表明，该 ELISA 可有效检测 BHV-1，可作为重要的诊断工具。
